Triazolone nicotinamide n-methyltransferase inhibitors and therapeutic uses thereof

Abstract

The present disclosure provides potent, selective, and orally bioavailable nicotinamide N-methyltransferase (NNMT) inhibitors with a triazolone-piperidine scaffold. The NNMT inhibitors are useful for treating tumors and cancers, metabolic disorders, and liver diseases.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 610682, filed December 15, 2023, which is incorporated herein by reference in its entirety.

[0003] Statement regarding government support

[0004] This invention was funded, in whole or in part, by the National Cancer Institute, the National Institutes of Health, the Union for Chemical Biology (Contract No. HHSN261200800001E, Protocol No. 16X129T10), and internal funding from the Early Translational Branch of the National Center for Translational Sciences. The government holds certain rights to this invention. Background Technology

[0005] Nicotinamide N-methyltransferase (NNMT) catalyzes the methylation of nicotinamide and similar compounds using the universal methyl donor S-adenosyl-L-methionine (SAM), producing 1-methylnicotinamide (1-MNA) and S-adenosyl-L-homocysteine ​​(SAH). Nicotinamide methylation via NNMT and SAM is the primary pathway for nicotinamide degradation, leading to the urinary excretion of 1-MNA. Nicotinamide is a biologically active form of vitamin B3 and is also known as oxidized nicotinamide adenine dinucleotide (NAD). + The precursor of NAD + Nicotinamide is a key molecule involved in the fundamental redox processes that regulate energy metabolism. Therefore, nicotinamide is an important cofactor that links cellular redox state with energy metabolism.

[0006] High expression of NNMT is a predictor of poor prognosis in many types of human cancer [Cao et al., Frontiers in Genetics, 13:1000515 (2022); and Dang et al., Open Medicine, 17:292-303 (2022)]. Elevated NNMT levels or activity promote cancer cell survival, growth, proliferation, migration, invasion and metastasis, as well as resistance to chemotherapeutic drugs [Li et al., Frontiers in Oncology, 12:894744 (2022); Pozzi et al., Biomolecules, 12:1173 (2022); and Eckert et al., Nature, 569:723-728 (2019)].

[0007] Furthermore, overexpression of NNMT in human adipose tissue is associated with obesity and insulin resistance [Pissios, Trends Endocrinol. Metab., 28(5):340-353 (2017)]. In addition, NNMT is upregulated in hepatic steatosis, hepatitis, liver fibrosis and cirrhosis [Liang et al., Genes & Diseases, 10:1883-1893 (2023)]. Summary of the Invention

[0008] This disclosure describes an effective, selective, and orally bioavailable nicotinamide N-methyltransferase (NNMT) inhibitor. The NNMT inhibitor has Formula I, or its tautomer or pharmaceutically acceptable salt:

[0009] I

[0010] Where X is an optionally substituted aryl or heteroaryl group, Y is an optionally substituted cycloalkyl, heterocyclic, aryl or heteroaryl group, and n is 1, 2 or 3.

[0011] Exemplary triazolone NNMT inhibitors reduce levels of 1-methylnicotinamide produced in vivo by NNMT (data not shown), reduce body weight in a high-fat diet-induced mouse obesity model (data not shown), and reduce tumor burden and metastasis in a mouse cancer model. Triazolone NNMT inhibitors may be used to treat tumors and cancers such as metabolic disorders of obesity and type 2 diabetes, as well as liver diseases. Brief description of the attached figures

[0012] The features and advantages of this disclosure will be better understood by referring to the following detailed description (which illustrates illustrative embodiments of this disclosure) and the accompanying drawings.

[0013] Figure 1 The results showed that intraperitoneal (ip) injection of ID8-luciferase ovarian cancer cells into C57BL / 6 mice, followed by intraperitoneal (ip) injections three times weekly of an NNMT inhibitor (NNMTi), a less active inferior compound (NNMTi-D), or a solvent control. Tumor burden was assessed using an in vivo imaging system (IVIS) (5–6 mice per group, univariate ANOVA). Treatment with compound 144 (NNMTi) significantly reduced tumor burden compared to treatment with the solvent or inferior compound 145 (NNMTi-D).

[0014] Figure 2The results showed that subcutaneous (sc) injection of MC38 colon cancer cells into C57BL / 6 mice (day 0) followed by intratumoral (it) injection of NNMTi, NNMTi-D, or a solvent control every 3 days after the tumor became palpable. Tumor growth was measured twice weekly (6–7 mice per group, two-factor ANOVA). Treatment with compound 144 (NNMTi) significantly reduced tumor volume compared to treatment with solvent or inferior compound 145 (NNMTi-D).

[0015] Figure 3A and Figure 3B The results showed that NNMT inhibition enhanced the therapeutic response to the immune checkpoint inhibitors PD-1 and CD47. Treatment with both compound 144 (NNMTi) and the PD-1 inhibitor monoclonal antibody (αPD1, InVivoMAb anti-mouse PD-1, clone RPM1-14, BioXCell, catalog number BE0146; RRID: AB_10949053) significantly and synergistically reduced tumor volume in a mouse subcutaneous MC38 tumor model. Figure 3A C57BL / 6 mice were subcutaneously injected with MC38 cells and treated with NNNMTi or NNNMTi-D via intratumoral (it) injection at the indicated time points, starting on day 5. αPD-1 treatment was administered via intraperitoneal injection on days 13, 16, and 21, highlighted with arrows (6–7 mice per group, two-factor ANOVA). Figure 3B C57BL / 6 mice were subcutaneously injected with MC38 cells and treated with intratumoral injection of compound 144 (NNMTi) or NNMTi-D starting on day 5, at the indicated time points. αCD47 is an antibody that blocks the CD47-SIRPα axis, increasing macrophage phagocytosis and leading to an antitumor effect. αCD47 antibodies were administered intraperitoneally on days 10, 15, 19, and 25, highlighted with arrows (7–8 mice per group, two-factor ANOVA).

[0016] Figure 4 The results showed that in a mouse model of low-grade serous ovarian cancer, treatment with compound 144 (NNMTi) significantly reduced tumor volume compared with treatment with solvent or inferior compound 145 (NNMTi-D).

[0017] Figure 5EO771-LMB breast cancer cells were injected intravenously (iv) into C57BL / 6 mice (day 0). From two days before injection (-2) to five days after injection, mice were treated daily by oropharyngeal inhalation (op) with NNMTi, NNMTi-D, or a solvent control. After 13 days, the lungs were fixed in formalin, embedded in paraffin, and the number and size of metastases were assessed by H&E staining (n=5 per group, single-factor ANOVA). Compound 144 (NNMTi) significantly reduced the size of metastases compared with treatment with solvent or inferior compound 145 (NNMTi-D).

[0018] Figure 6 EO771-LMB cells were injected intravenously (iv) into C57BL / 6 mice (day 0). Mice were treated daily with NNMTi or NNMTi-D from two days before (-2) to five days after injection. αPD-1 was administered intraperitoneally (ip) on days 7 and 11 as indicated by arrows. Lung metastases were assessed by H&E staining after 15 days (6–8 mice per group, single-factor ANOVA) (prevention study). The combination of compound 144 (NNMTi) and αPD-1 antibody significantly reduced metastatic lesion size compared with αPD-1 antibody treatment alone.

[0019] Figure 7A and Figure 7B The results showed that whole-body Nnmt- / - mice exhibited reduced tumor burden. Figure 7A Nnmt+ / + (n=6) and Nnmt- / - (n=6) mice were injected into the left ovarian cyst and injected with the mouse HGS-2 ovarian cancer cell line. The tumor burden of primary ovarian and omental tumors was assessed after 42 days (Student's t-test). Figure 7B Nnmt+ / + (n=6) and Nnmt- / - (n=6) mice were injected into the left ovarian cyst and injected with the mouse HGS-3 ovarian cancer cell line. The tumor burden of primary ovarian and omental tumors was assessed after 42 days (Student's t-test).

[0020] Figure 8 The results showed that whole-body Nnmt- / - mice exhibited reduced tumor burden. Nnmt-knockout in the tumor microenvironment reduced tumor burden. Nnmt+ / + wild-type (n=10) and Nnmt- / - knockout mice (n=11) were injected into the mammary fat pads, followed by injection of mouse EO771-LMB breast cancer cell line, and tumor growth was monitored twice weekly by caliper measurement (one-way ANOVA).

[0021] Figure 9A and Figure 9B The results showed that NNMT inhibition significantly reduced tumor burden and metastasis in an orthotopic mouse model of ovarian cancer. Figure 9AC57Bl6 mice were injected with the HGS-2 ovarian cancer cell line into their left ovarian cysts, and starting 4 days after the cancer cell injection, they were treated daily by intraperitoneal injection of compound 144 (NNMTi) at 25 mg / kg for 5 weeks (formulation: 10% NMP, 20% PEG400, 70% Solutol). Figure 9B C57Bl6 mice were injected with BPNM ovarian cancer cells into the left ovarian cyst, and starting 4 days after cancer cell injection, they were treated daily for 12 days with compound 144 (NNMTi) at 25 mg / kg via intraperitoneal injection (formulation: 10% NMP, 20% PEG400, 70% Solutol). BPNM (Trp53− / −R172HBrca1− / −Pten− / −Nf1− / −MycOE genotype) was HR-deficient (S. Iyer Cancer Discovery 2021).

[0022] Figure 10 The results showed that NNMT inhibition significantly reduced tumor burden in an orthotopic mouse model of breast cancer. C57Bl6 mice were injected with the mouse EO771-LMB breast cancer cell line into the mammary fat pad, and starting 4 days after cancer cell injection, mice were treated daily by intraperitoneal injection of compound 144 (NNMTi) (25 mg / kg) or a solvent control. (Formulation: 10% NMP, 20% PEG400, 70% Solutol) (One-factor ANOVA).

[0023] Figure 11 The results showed that NNMT inhibition improved the therapeutic response to local radiotherapy. C57BL / 6 mice were subcutaneously injected with LLC (Lewis lung cancer) cells and treated daily from day 4 by intraperitoneal injection of either the solvent or compound 144 (NNMTi). On day 13, the tumors were locally irradiated with 20 Gy (10 mice per group, two-factor ANOVA).

[0024] Figure 12A , Figure 12B and Figure 12C This demonstrates the effect of NNMT inhibition on CAF in vivo. Figure 12AEffects of NNMT inhibition on CAF in vivo in a breast cancer model. NNMT inhibition reduced the abundance of established CAF subtypes in vivo. Flow cytometry: C57Bl6 mice were injected with the mouse EO771-LMB breast cancer cell line into the mammary fat pad. Four days after injection, they were treated daily by intraperitoneal injection of compound 144 (NNMTi) (25 mg / kg) or a solvent control. Eighteen days after cancer cell injection, the tumors were dissected and flow cytometry was performed. Ly6C+ inflammatory CAF (iCAF), α-SMA+ myofibroblast CAF (myCAF), MHCII+ antigen-presenting CAF (apCAF), and PDGFRα+ vascular CAF (vCAF) were identified. Figure 12B In an orthotopic ovarian cancer model, compound 144 (NNMTi) treatment reduced the abundance of Ly6C+iCAF and MHCII+myCAF. Flow cytometry (gated): C57Bl6 mice were injected with the mouse HGS-2 ovarian cancer cell line into their ovarian cysts. Four days post-injection, they were treated daily via intraperitoneal injection with either compound 144 (NNMTi) (25 mg / kg) or a solvent control. Nineteen days post-injection, the tumors were dissected and subjected to flow cytometry. Figure 12C Compound 144 (NNMTi) treatment increased H3K27 trimethylation in CAF in vivo. Flow cytometry: Absolute quantification of low H3K27me expression in CD45-EPCAM-CD31-Podoplanin+ CAF in primary EO771-LMB tumors treated with compound 144 (NNMTi) or solvent control.

[0025] Figure 13 The effect of NNMT inhibition on complement factor C3 abundance was shown. Treatment with compound 144 (NNMTi) reduced the secretion of complement factors in vivo. Enzyme-linked immunosorbent assay (ELISA): EO771-LMB primary tumor homogenates treated with compound 144 (NNMTi) or solvent control were homogenized 18 days after cancer cell injection, and complement factor C3 protein was detected (Student's t-test).

[0026] Figure 14A , Figure 14B and Figure 14C This study demonstrates the effect of compound 144 (NNMTi) treatment on tumor immune cell infiltration. Compound 144 (NNMTi) treatment reduced the abundance of immunosuppressive monocytes of myeloid origin (M-MDSCs; high Ly6C, high PD-L1) and increased CD8+ T cell responses. Figure 14AMC38 colon cancer cells were subcutaneously (sc) injected into C57BL / 6 mice (day 0). Five days later, once the tumor was palpable, mice were treated every 3 days with either compound 144 (NNMTi) or NNNMTi-D as a control by intratumoral injection (it). Fifteen days after cancer cell injection, MC38 tumors were dissected and subjected to 19-color flow cytometry (6-8 mice per group, Student's t-test). Figure 14B Fifteen days after injection of cancer cells, MC38 tumors were dissociated, stimulated with PMA / ionomycin, and the production of effector cytokines was quantified (Student's t-test). Figure 14C C57Bl6 mice were orthotopically injected with HGS-2 ovarian cancer cells or EO771-LMB breast cancer cells. Four days after cancer cell injection, mice were treated daily with compound 144 (NNMTi) via intraperitoneal injection at a dose of 25 mg / kg. Eighteen days after cancer cell injection, primary tumors were dissected and analyzed by flow cytometry (Student's t-test).

[0027] Figure 15A and Figure 15B This demonstrates the effect of compound 144 (NNMTi) treatment on the tumor metabolome. Figure 15A Experimental plan: MC38 cells were subcutaneously injected into C57BL / 6 mice. Thirteen days later, the tumors were treated with solvent, NNNMTi-D, or compound 144 (NNMTi) via intratumoral injection. Tumors were then sampled at different time points after injection. Figure 15B Tumor samples were extracted for liquid chromatography-mass spectrometry analysis. Core metabolites of the methionine and nicotinamide cycles were shown to demonstrate the effect of NNMT inhibition on the levels of these metabolites (n=5 mice / group. Conventional one-way ANOVA).

[0028] Figure 16 This demonstrates that NNMT inhibition exerts its tumor-reducing effect by targeting the TME, rather than by altering cancer cell growth. To determine whether NNMT inhibition in cancer cells reduces tumor growth in vivo, we injected MC38 colon cancer cells into systemically Nnmt- / - mice and administered either the solvent or compound 144 (NNMTi). These mice were completely lacking in NNMT, so treatment with compound 144 (NNMTi) should have no specific effect on TME-mediated cells. Since the cancer cells were Nnmt-wildtype and expressed NNMT, any effect of NNMT inhibition on cancer cell growth would be detectable. However, we did not observe tumor reduction in our experiments. Invention Details

[0029] General Public Content

[0030] Although various embodiments of this disclosure are described herein, those skilled in the art will understand that these embodiments are provided by way of example only. Many modifications and changes, variations and substitutions, to the embodiments described herein will be apparent to those skilled in the art without departing from this disclosure. It should be understood that various alternatives to the embodiments described herein may be employed in implementing this disclosure. It should also be understood that each embodiment of this disclosure may be optionally combined with one or more other embodiments described herein that are consistent with that embodiment.

[0031] When a combination is disclosed, it should be understood that every possible sub-combination of the elements in that combination is also disclosed. Conversely, when different elements or groups of elements are disclosed individually, their combinations are also disclosed.

[0032] When elements are presented in a list format or as alternative members of a group (e.g., a Markush group), it should be understood that each possible subgroup of elements is also exposed, and any one or more elements can be removed from the list or group.

[0033] When describing a range of values, it should be understood that the endpoints of the range, each intermediate integer value between the endpoints, each fraction thereof, and each subrange are specifically disclosed. All endpoints of the range are included within the range and can be independently combined. If the value has an inherent limit, that inherent limit is specifically disclosed. If a value is explicitly described, it should be understood that values ​​approximately the same as the described value are specifically disclosed.

[0034] It should also be understood that, unless explicitly stated otherwise, in any method described or claimed herein that includes more than one action or step, the order of the actions or steps of the method is not necessarily limited to the order in which the actions or steps of the method are described, but this disclosure covers embodiments where the order is so limited.

[0035] To be further understood, generally, where an embodiment is referred to as containing one or more features in the specification or claims, this disclosure also covers embodiments consisting of or substantially consisting of said features.

[0036] It should also be understood that, regardless of whether such specific exclusion is described in the specification, any embodiment of this disclosure, such as any embodiment or compound found in the prior art, may be expressly excluded from the claims.

[0037] Furthermore, it should be understood that any functional language used in any claim should not be interpreted as the “apparatus plus function” language under 35 USC §112(f) unless it is expressly indicated in the claim by using the terms “apparatus for…” or “step for…”.

[0038] To be further understood, this disclosure covers all analogues, derivatives, prodrugs, metabolites, salts, solvates, hydrates, cages, and polymorphs of the compounds / substances disclosed herein, as appropriate. In certain instances of this disclosure, the specific use of the terms "analyte," "derivative," "prodrug," "metabolite," "salt," "solvate," "hydrate," "cage," or "polymorph" with respect to a compound / substance or group of compounds / substances should not be construed as an intention to omit any of these forms in other instances where the compound / substance or group of compounds / substances is mentioned or shown without recounting any of these forms, unless otherwise stated or clearly indicated by the context.

[0039] It should also be understood that this disclosure covers all possible tautomers, all possible regioisomers, and all possible stereoisomers, including two enantiomers in substantially pure forms and all possible diastereomers, as appropriate, and mixtures of enantiomers (including racemic mixtures) and mixtures of two or more diastereomers in any proportion, not just the specific tautomers, regioisomers, and stereoisomers indicated by the drawn structures or names. Some embodiments of this disclosure relate to specific tautomers, regioisomers, and stereoisomers indicated by the drawn structures or names. In certain instances of this disclosure, the specific descriptive phrases “or its tautomers,” “or its regioisomers,” “or its stereoisomers,” etc., relating to a compound / substance or a group of compounds / substances should not be construed as an intention to omit other possible tautomers, regioisomers, and stereoisomers of the compound / substance or the group of compounds / substances in other instances of this disclosure, where the term “compound,” etc., is used, or the compound / substance or the group of compounds / substances is mentioned or shown without the phrases “or its tautomers,” “or its regioisomers,” “or its stereoisomers,” etc., unless otherwise stated or clearly indicated by the context.

[0040] Headings are included in this document for reference and to help locate certain sections. Headings are not intended to limit the scope of the embodiments and concepts described in the sections under a heading, and these embodiments and concepts are applicable to other parts of the disclosure.

[0041] All patent and non-patent documents cited in this article are incorporated herein by reference in their entirety, as if each patent or non-patent document were specifically and individually indicated to be incorporated herein by reference in its entirety.

[0042] definition

[0043] Unless otherwise defined herein or expressly indicated by its use, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0044] As used in the specification and claims, the indefinite articles “a” and “an” and the definite article “the” can include both plural and singular references, unless otherwise expressly stated or indicated by the context.

[0045] The terms “or / and” and “and / or” mean “either…, or…, or both… and…” when referring to two elements, and “either…, or…, or any combination or all of them” when referring to three or more elements. For example, the phrase “A or / and B” means “either A, or B, or both A and B”, and the phrase “A, B or / and C” means “either A, B, or C, or any combination or all of them”.

[0046] As used in the specification and claims, all transitional phrases such as “comprising,” “containing,” “having,” “including,” “owning,” “holding,” “carrying,” “with,” “consisting of,” “characterized in,” etc., are open-ended and inclusive, meaning that they include, but are not limited to, and do not exclude additional, unlisted elements or method steps. Only the transitional phrase “consisting of” is closed-ended, meaning that it excludes any additional, unlisted elements or method steps, while the transitional phrase “substantially constitutes” is semi-closed-ended, meaning that it only allows the inclusion of additional, unlisted elements or method steps that do not materially affect the basic and novel features of that particular embodiment.

[0047] The term “exemplary” as used herein means “serving as an example, instance, or illustration.” Any implementation or feature described herein as “exemplary” should not be construed as preferred or advantageous over other implementations or features.

[0048] The terms “about” or “approximately” mean an acceptable error for a particular value as determined by one of ordinary skill in the art, depending in part on how the value was measured or determined. In some embodiments, the terms “about” or “approximately” mean within one standard deviation. In some embodiments, when no specific error range is stated (e.g., the standard deviation of the average given in a chart or data table), the terms “about” or “approximately” mean the range that covers the stated value and the range included by rounding up or down to the stated value, taking into account significant figures. In some embodiments, the terms “about” or “approximately” mean within ±10% or ±5% of the specified value. Whenever the terms “about” or “approximately” appear before the first value in a series of two or more values ​​or before the first range of values ​​in a series of two or more values, the terms “about” or “approximately” apply to each value in the series or each range of values ​​in the series.

[0049] In some embodiments, the term "substantially all" means at least about 90%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the term "substantially none" means no more than about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% by weight or molar concentration, or no more than about 1000 ppm, about 500 ppm, about 400 ppm, about 300 ppm, about 200 ppm, or about 100 ppm.

[0050] In some embodiments, the term "substantially pure" means at least about 90%, about 95%, about 96%, 97%, about 98%, or about 99% pure. In some embodiments, the term "substantially stereochemically pure" means at least about 80% enantiomer excess (90% of one enantiomer to 10% of the other enantiomer) or 90% enantiomer excess (95% of one enantiomer to 5% of the other enantiomer), or at least about 80% diastereomer excess (90% of one diastereomer to 10% of all other diastereomers) or at least about 90% diastereomer excess (95% of one diastereomer to 5% of all other diastereomers).

[0051] Whenever the term “at least” or “greater than” appears before the first value in a series of two or more values, the term “at least” or “greater than” applies to each value in the series.

[0052] Whenever the term “not more than” or “less than” appears before the first value in a series of two or more values, the term “not more than” or “less than” applies to each value in the series.

[0053] The term "pharmaceuticalally acceptable" means that a substance (e.g., an active ingredient or excipient) is generally safe, non-toxic, and suitable for use in contact with the cells, tissues, and organs of a target without excessive irritation, allergic reactions, immunogenicity, or other adverse effects. "Pharmaceuticalally acceptable" excipients or carriers in a pharmaceutical composition are also compatible with the other components of the composition.

[0054] The term "therapeutic effective amount" refers to an amount of compound, when administered to a subject or in ex vivo, sufficient to prevent, reduce the risk of development, delay onset, slow progression, or cause remission of the treated medical condition, or at least partially alleviate or improve the medical condition or one or more symptoms or complications thereof, at least in a portion of the subject taking the compound or receiving ex vivo treatment with the compound. The term "therapeutic effective amount" also refers to an amount of compound sufficient to elicit a biological or medical response in cells, tissues, organs, systems, animals, or humans sought by researchers, veterinarians, physicians, or clinicians.

[0055] The term "treatment" includes alleviating, improving, reducing the incidence, frequency, or severity of a medical condition, slowing or stopping its progression, reversing or eliminating a medical condition or one or more symptoms or complications associated with it, and alleviating, improving, or eradicating one or more causes of the condition. References to "treatment" of a medical condition include prevention of the condition. The term "prevention" includes eliminating, reducing the risk or likelihood of, and delaying the onset of a medical condition or one or more symptoms or complications associated with it.

[0056] The term "medical condition" (or simply "condition") includes both disease and disorder. The terms "disease" and "disorder" are used interchangeably in this document.

[0057] The term “object” refers to an animal, including but not limited to mammals such as primates (e.g., humans, chimpanzees, or monkeys), rodents (e.g., rats, mice, guinea pigs, gerbils, or hamsters), lagomorphs (e.g., rabbits), bovids (e.g., cattle), suidae (e.g., pigs), capridae (e.g., sheep), equines (e.g., horses), canids (e.g., dogs), or felines (e.g., cats). When referring to an object / patient suffering from a medical condition (e.g., mammalian object / patient, such as human object / patient), the terms “object” and “patient” may be used interchangeably herein.

[0058] For example, a "regulator" of a receptor or enzyme can be an activator or inhibitor of that receptor or enzyme, and can increase or decrease the activity and / or level of that receptor or enzyme.

[0059] The term "compound" or similar terms (e.g., "molecule") encompasses a salt, solvate, hydrate, cage compound, and polymorph of the compound, or a salt of the compound. A "solvate" of a compound comprises stoichiometric or non-stoichiometric solvent molecules (e.g., water, acetone, or an alcohol [e.g., ethanol]) non-covalently bound to the compound. A "hydrate" of a compound comprises stoichiometric or non-stoichiometric water molecules non-covalently bound to the compound. A "cage compound" of a compound comprises molecules of a substance (e.g., a solvent) embedded in the crystal structure of the compound. A "polymorph" of a compound is the crystalline form of the compound. In certain instances of this disclosure, the specific use of the terms "salt," "solvate," "hydrate," "cage compound," or "polymorph" with respect to a compound or group of compounds should not be construed as an intention to omit any of these forms in other instances of this disclosure, where the term "compound" or similar terms (e.g., "molecule") are used, or where the compound or group of compounds is mentioned or shown without recounting any of these forms, unless otherwise stated or clearly indicated by the context.

[0060] The terms “halogen,” “halide,” and “halo” refer to fluorine, chlorine, bromine, and iodine.

[0061] The term "alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group, wherein the alkyl group may optionally be substituted by one or more substituents as described herein. In some embodiments, the alkyl group is having a carbon number of 1 to 20 (C0). 1-20 ), 1 to 10 (C 1-10 ) or 1 to 6 (C 1-6 A straight-chain saturated monovalent hydrocarbon group with 3 to 20 carbon atoms, or a group with 3 to 20 carbon atoms. 3-20 ), 3 to 10 (C 3-10 ) or 3 to 6 (C 3-6 A branched, monovalent hydrocarbon group consisting of 1 carbon atom. For example, the term "C 1-6 "Alkyl" refers to a straight-chain saturated monovalent hydrocarbon group having 1 to 6 carbon atoms or a branched saturated monovalent hydrocarbon group having 3 to 6 carbon atoms. Straight-chain C 1-6 and branch chain C 3-6 Alkyl groups are also referred to as "lower alkyl groups". Non-limiting examples of alkyl groups include methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including all isomers such as n-butyl, isobutyl, sec-butyl and tert-butyl), pentyl (including all isomers such as n-pentyl and isopentyl) and hexyl (including all isomers such as n-hexyl).

[0062] The terms “alkylene” and “-alkyl-” refer to divalent alkyl groups, which may optionally be substituted by one or more substituents as described herein.

[0063] The term "haloalkyl" refers to an alkyl group substituted with one or more halogen atoms. A haloalkyl group may optionally be substituted with one or more additional substituents as described herein. Examples of haloalkyl groups include, but are not limited to, fluoroalkyl groups, such as -CH2F, -CHF2, and -(CH2). n CF3, and perfluoroalkyl groups, such as -CF3 and -(CF2). n CF3, where n is 1, 2, 3, 4 or 5.

[0064] The term "heteroalkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group containing one or more heteroatoms independently selected from O, N, and S. In some embodiments, one or more heteroatoms are located in the backbone of the straight-chain or branched hydrocarbon group. The terms "heteroalkylene" and "-heteroalkyl-" refer to divalent heteroalkyl groups. Heteroalkyl and -heteroalkyl- groups may optionally be substituted with one or more substituents as described herein. Examples of heteroalkyl and -heteroalkyl- groups include, but are not limited to, -(CH2). m -(O or S)-(CH2) n CH3 and -(CH2) m -(O or S)-(CH2) p - where m is 1, 2 or 3, n is 0, 1 or 2, and p is 1, 2 or 3.

[0065] The term "alkoxy" refers to an -O-alkyl group, which may optionally be substituted with one or more substituents as described herein.

[0066] The term “haloalkoxy” refers to an O-haloalkyl group, which may optionally be substituted with one or more substituents as described herein.

[0067] Examples of -O-heteroalkyl and -O-heteroalkyl- groups include, but are not limited to, ethylene glycol groups and polyethylene glycol (PEG) groups, including, but not limited to, -(OCH2CH2). n -OR and -(OCH2CH2) n -O-, where R is hydrogen or alkyl, and n is 1, 2, or 3. The -O-heteroalkyl and -O-heteroalkyl- groups may optionally be substituted by one or more substituents as described herein.

[0068] The term "cycloalkyl" refers to a cyclic, saturated or partially unsaturated, bridged or unbridged monovalent hydrocarbon group, which may optionally be substituted with one or more substituents as described herein. In some embodiments, the cycloalkyl group has a 3 to 10 (C) 3-10 ), or 3 to 8 (C 3-8 ), or 3 to 6 (C 3-6( ) carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, decahydronaphthyl, and adamantyl. The term "-cycloalkyl-" refers to a divalent cycloalkyl group, which may optionally be substituted with one or more substituents as described herein.

[0069] The terms "heterocyclic group" and "heterocyclic" refer to a monocyclic non-aromatic group or a polycyclic group containing at least one non-aromatic ring, wherein the at least one non-aromatic ring contains one or more heteroatoms independently selected from O, N, and S. The non-aromatic ring containing one or more heteroatoms may be attached to or fused to one or more saturated, partially unsaturated, or aromatic rings, which may contain only cyclic carbon atoms or may contain one or more cyclic heteroatoms. In some embodiments, the heterocyclic group or heterocyclic group has 3 to 15, 3 to 12, 3 to 10, 3 to 8, or 3 to 6 ring atoms. In some embodiments, the heterocyclic group or heterocyclic group is a monocyclic, bicyclic, or tricyclic system, which may include fused or bridged ring systems, and wherein nitrogen or sulfur atoms may optionally be oxidized, nitrogen atoms may optionally be quaternized, and one or more rings may be fully or partially saturated, or aromatic. Heterocyclic groups or heterocyclic compounds can be attached to any heteroatom or carbon atom in the host structure to produce stable compounds. Examples of heterocyclic groups or heterocyclic compounds include, but are not limited to, azirroheptanyl, cyclohexylimino, azirroheptanyl, azirroheptanyl, azirroheptanyl, azirroheptanyl, benzodioxane (e.g., 1,4-benzodioxane), methylenedioxyphenyl (e.g., 1,3-methylenedioxyphenyl), benzofuranone, benzopyranone, benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothiophene, benzothiopyranyl, β-carbolinyl, chromanyl, decahydroisoquinolinyl, dihydrobenzisothiazinyl, dihydrobenzisothiazinyl, dihydrofuranyl, dihydropyranyl, dihydropyrazinyl, dihydropyridyl, dihydropyrimidinyl, dihydropyrrolyl, dioxolaneyl, dioxaneyl, dioxaneyl, furanyl, dioxane ... The following groups are included in the class of heterocyclic compounds: ketone, imidazolinyl, imidazolinyl, indololinyl, indolazinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothiophenyl, isochoryl, isoindololinyl, isothiazolyl, isoxazolyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinone, oxazolyl, oxehappyl, oxehappyl, oxehappylylene, oxehappylbutyl, ethylene oxide, oxehappyloctyl, piperazinyl, piperidinyl, 4-piperidinoneyl, pyrrolyl, pyrrololinyl, quininecycloyl, tetrahydrofuranyl, tetrahydrofuranyl (oxehappyl), tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothiophenyl (tetrahydrothiophenyl or thiohexacyclopentyl), thiomorpholinyl (thiomorpholinyl), thiazolinyl, and 1,3,5-trithiaalkyl. The term "-heterocyclic-" refers to a divalent heterocyclic group. Heterocyclic groups or heterocyclic groups, as well as -heterocyclic- groups, may optionally be substituted with one or more substituents as described herein.

[0070] The term "aryl" refers to a monocyclic aromatic hydrocarbon group or a polycyclic group containing at least one aromatic hydrocarbon ring. In some embodiments, the aryl group has 6 to 15, 6 to 12, or 6 to 10 ring atoms. Non-limiting examples of aryl groups include phenyl, naphthyl, fluorenyl, azulel, anthracenel, phenanthryl, biphenyl, and terphenyl. The aromatic hydrocarbon ring of the aryl group may be attached to or fused to one or more saturated, partially unsaturated, or aromatic rings, which may contain only cyclic carbon atoms (e.g., biphenyl, dihydronaphthyl, indole, dihydroindole, and tetrahydronaphthyl) or may contain one or more cyclic heteroatoms (e.g., indololinyl, isoindolinyl, 2,3-dihydrobenzofuranyl, 1,3-methylenedioxyphenyl, chromium, and 1,4-benzodioxane). The term "-aryl-" refers to a divalent aryl group. The aryl and -aryl- groups may optionally be substituted by one or more substituents as described herein.

[0071] The term "heteroaryl" refers to a monocyclic aromatic group or a polycyclic group containing at least one aromatic ring, wherein the at least one aromatic ring contains one or more heteroatoms independently selected from O, N, and S. The heteroaryl ring may be attached to or fused to one or more saturated, partially unsaturated, or aromatic rings, which may contain only cyclic carbon atoms or may contain one or more cyclic heteroatoms. The heteroaryl group may be attached to the host structure at any heteroatom or carbon atom that produces a stable compound. In some embodiments, the heteroaryl group has 5 to 15, 5 to 12, or 5 to 10 ring atoms. Examples of monocyclic heteroaryl groups include, but are not limited to, pyrroleyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thiophenyl, oxadiazolyl, triazolyl, tetrazolyl, pyridinyl, pyridoneyl, pyrazinyl, pyrimidinyl, pyridazinoneyl, and triazinyl. Non-limiting examples of bicyclic heteroaryl groups include indolyl, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, benzoxazolyl, benzoisoxazolyl, 1,2,3-benzoxadiazolyl, 2,1,3-benzoxadiazolyl (benzofuranyl), benzothiophene, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzoimidazolyl, benzotriazolyl, indolazinyl, benzofuranyl, isobenzofuranyl, crononeyl, coumarinyl, cinolinyl, quinazolinyl, quinoxalinyl, indolyl, naphthidyl, phthalazinyl, quinazolinyl, purinyl, pyrrolopyridyl, furanopyridyl, thienopyridyl, dihydroisoindolyl, and tetrahydroquinolinyl. Examples of tricyclic heteroaryl groups include, but are not limited to, carbazolyl, benzoindolyl, dibenzofuranyl, phenanthrolinel, acridinel, phenanthidyl, xanthonyl, and phenothiazinyl. The term "-heteroaryl-" refers to a divalent heteroaryl group. Heteroaryl and -heteroaryl- groups may optionally be substituted with one or more substituents as described herein.

[0072] The terms “-alkylcycloalkyl,” “-alkylheterocyclic,” “-alkylaryl,” and “-alkylheteroaryl” refer to an alkyl group that is substituted by one or more cycloalkyl groups, one or more heterocyclic groups, one or more aryl groups, or one or more heteroaryl groups, respectively. The -alkylcycloalkyl, -alkylheterocyclic, -alkylaryl, or -alkylheteroaryl groups may optionally be substituted by one or more additional substituents as described herein.

[0073] Each group described herein (including, but not limited to, monovalent and divalent alkyl, haloalkyl, monovalent and divalent heteroalkyl, -O-alkyl, -O-haloalkyl, -O-heteroalkyl, monovalent and divalent cycloalkyl, monovalent and divalent heterocyclic, monovalent and divalent aryl, monovalent and divalent heteroaryl, -alkylcycloalkyl, -alkylheterocyclic, -alkylaryl, and -alkylheteroaryl), whether as a base group or as a substituent, may optionally be substituted by one or more substituents. In some embodiments, each group described herein may optionally be substituted by one, two, three, four, five, or six substituents independently selected from: halides, cyano (-CN), nitro (-NO2), hydroxyl (-OH), mercapto (-SH), amino (-NH2), -OR 11 -SR 11 -NR 12 R 13 -C(=O)R 11 -C(=O)OR 11 -OC(=O)R 11 -C(=O)NR 12 R 13 -NR 12 C(=O)R 11 -OC(=O)OR 11 -OC(=O)NR 12 R 13 -NR 12 C(=O)OR 11 -NR 11 C(=O)NR 12 R 13 Alkyl, haloalkyl, alkoxy, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein:

[0074] R 11 Each time it appears, it is independently hydrogen, alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, -alkylcycloalkyl, -alkylheterocyclic, -alkylaryl, or -alkylheteroaryl; and

[0075] R 12 and R 13Each time it appears, it is independently hydrogen, alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, -alkylcycloalkyl, -alkylheterocyclic, -alkylaryl or -alkylheteroaryl, or R. 12 and R 13 And the nitrogen atoms they are connected to form heterocycles or heteroaromatic rings.

[0076] NNMT inhibitors

[0077] This disclosure provides an inhibitor of nicotinamide N-methyltransferase (NNMT) having Formula I, or its tautomer, pharmaceutically acceptable salt, solvate, hydrate, cage, or polymorph:

[0078] I

[0079] in:

[0080] X is an aryl or heteroaryl group, wherein the aryl or heteroaryl group may optionally have one or more (e.g., two or three) substituents independently selected from the following: halide, cyano, nitro, C 1-6 Alkyl, -CF3, -OH, -OR 1 -NH2, -NR 2 R 3 -(C=O)NR 4 R 5 and the proteolytic targeting portion (e.g., a thalidomide-based proteolytic targeting portion), and wherein the C 1-6 Alkyl groups may optionally be replaced by -OH or -OR 1 -NH2 or -NR 2 R 3 replace;

[0081] Y is a cycloalkyl, heterocyclic, aryl, or heteroaryl group, wherein the cycloalkyl, heterocyclic, aryl, or heteroaryl group may optionally have one or more (e.g., two or three) substituents independently selected from the following: halide, cyano, nitro, C 1-6 Alkyl, -CF3, -OH, -OR 1 -NH2, -NR 2 R 3 and -(C=O)NR 4 R 5 And wherein C 1-6 Alkyl groups may optionally be replaced by -OH or -OR 1 -NH2 or -NR 2 R 3 replace;

[0082] R 1 It is hydrogen, C 1-6Alkyl, -CHF2, -CF3 or C 3-6 Cycloalkyl, wherein the alkyl or cycloalkyl group may optionally be converted to -OH, C 1-4 Alkoxy, -NH2 or -NR 2 R 3 replace;

[0083] R 2 and R 3 Independently hydrogen, C 1-6 Alkyl or -C(=O)-(C 1-6 Alkyl), or R 2 and R 3 And the nitrogen atoms they are attached to form 3 to 6-membered heterocycles, or 5-membered heteroaromatic rings, or 6-membered heteroaromatic rings;

[0084] R 4 and R 5 Independently hydrogen or C 1-6 alkyl;

[0085] n is 1, 2, or 3; and

[0086] The stereocenter marked with an asterisk (*) may have (S)-stereochemistry or (R)-stereochemistry or may be racemic;

[0087] The conditions are:

[0088] X is not an unsubstituted phenyl or 4-chlorophenyl; and

[0089] The compound of formula I is not:

[0090] , ,

[0091] or ,

[0092] Or its tautomers or salts.

[0093] In some embodiments, X is an optionally substituted 5- to 10-membered monocyclic or bicyclic aryl or heteroaryl group. In further embodiments, X is selected from 2-CN-phenyl, 3-CN-phenyl, 3-CH3-phenyl, 3-OCH3-phenyl, 3-(C(=O)NH2)-phenyl, 4-Br-phenyl, 4-OCH3-phenyl, 4-OCHF2-phenyl, 4-(morpholino-N-yl)-phenyl, 3-CN-4-CH3-phenyl, 3-F-4-OCH3-phenyl, 3-F-5-CH3-phenyl, 3,5- Dimethyl-phenyl, 1-naphthyl, N-CH3-pyrazol-4-yl, 4-CH3-pyridin-2-yl, 5-CN-pyridin-2-yl, 5-CH3-pyridin-2-yl, 5-CF3-pyridin-2-yl, 6-CH3-pyridin-2-yl, 2-CH3-pyridin-3-yl, 4-CH3-pyridin-3-yl, 5-CH3-pyridin-3-yl, 6-CH3-pyridin-3-yl, 2-CH3-pyridin-4-yl

[0094] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0095] In some embodiments, the aryl or heteroaryl X group is replaced by or linked to a proteolytic targeting moiety. In some embodiments, the proteolytic targeting moiety is a thalidomide-based proteolytic targeting moiety. The linking of the triazolone compound to the proteolytic targeting moiety forms a proteolytic targeting chimera (PROTAC), which induces selective intracellular proteolytic digestion of NNMT. The proteolytic targeting moiety (e.g., thalidomide) binds to an E3 ubiquitin ligase, while the triazolone compound selectively binds to NNMT. Recruitment of the E3 ligase to NNMT leads to ubiquitination, followed by proteasome degradation of NNMT.

[0096] In some implementation schemes, regarding Y:

[0097] The cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, or cyclohexenyl;

[0098] The heterocyclic group is tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, piperazineyl, or morpholinyl;

[0099] The aryl group is phenyl, naphthyl, dihydroisobenzofuranyl, methylenedioxyphenyl, benzodioxane, isoindolinyl, 1,2,3,4-tetrahydroquinolinyl, or 1,2,3,4-tetrahydroisoquinolinyl; and

[0100] The heteroaryl group is furanyl, thiophene, pyrrole, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, quinolinyl, or isoquinolinyl;

[0101] The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted as defined above.

[0102] In a further embodiment, Y is selected from cyclohexyl, 1,1-diF-cyclohexyl-4-yl, 2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl, piperidin-3-yl, N-CH3-piperidin-3-yl, N-acetyl-piperidin-3-yl, piperidin-4-yl, N-CH3-piperidin-4-yl, N-acetyl-piperidin-4-yl, phenyl, 2-F-phenyl, 2-Cl-phenyl, 2-CF3-phenyl, 2-OCH3-phenyl, 2-OCH2CH3-phenyl, 2-OCH2CH2CH3-phenyl, 3-F-phenyl, 3-Cl-phenyl, 3-CH3-phenyl, 3-CF3-phenyl, 3-OCH3-phenyl, 3-(OCH2CH2OH)-phenyl 3-(piperidin-N-yl)-phenyl, 3-(morpholin-N-yl)-phenyl, 3-(imidazol-N-yl)-phenyl, 4-F-phenyl, 4-Cl-phenyl, 4-Br-phenyl, 4-CH3-phenyl, 4-ethyl-phenyl, 4-isopropyl-phenyl, 4-CH2OH-phenyl, 4-CF3-phenyl, 4-OH-phenyl, 4-OCH3-phenyl, 4-OCHF2-phenyl, 4-OCF3-phenyl, 4-(OCH2CH2OCH3)-phenyl, 4-(O-cyclopentyl)-phenyl, 4-N(CH3)2-phenyl, 4-(N-acetyl)-phenyl, 4-(imidazol-N-yl)-phenyl, 4-(pyrazol-N-yl)-phenyl, 2-CH3-3-F-phenyl 2-OCH3-3-F-phenyl, 2-F-4-Cl-phenyl, 2,4-diCl-phenyl, 2,4-dimethyl-phenyl, 2-F-4-methoxy-phenyl, 2-F-4-ethoxy-phenyl, 2-OH-4-OCH3-phenyl, 3-F-4-Cl-phenyl, 3,4-diCl-phenyl, 3-CH3-4-F-phenyl, 3-CF3-4-CH3-phenyl, 3,5-dimethyl-phenyl, 1-naphthyl, 2-naphthyl, 1,2,3,4-tetrahydroisoquinoline-8-yl, 4-isoindolinyl, 5-dihydroisobenzofuranyl, 1,3-methylenedioxyphenyl-5-yl, 1,4-benzodioxane-6-yl, N-CH3-pyrazol-4-yl, N-CH3- 3-CH3-pyrazol-4-yl, N-methyl-imidazol-2-yl, N-ethyl-imidazol-2-yl, 5-CH3-isoxazol-3-yl, 2-pyridyl, 3-F-pyridyl-2-yl, 3-CH3-pyridyl-2-yl, 4-Cl-pyridyl-2-yl, 4-CH3-pyridyl-2-yl, 4-OCH3-pyridyl-2-yl, 5-Cl-pyridyl-2-yl, 6-CH3-pyridyl-2-yl, 4,6-dimethyl-pyridyl-2-yl, 3-pyridyl, 2-OCH3-pyridyl-3-yl, 6-OCH3-pyridyl-3-yl, 4,6-diCl-pyridyl-3-yl, 4-CH3-pyrimidin-2-yl, 5-CH3-pyrimidin-2-yl, 2-quinolinyl, and 5-isoquinolinyl.

[0103] In some implementations, n is 2. In other implementations, n is 1.

[0104] In some embodiments, the compound of formula I is selected from the following compounds, their tautomers, pharmaceutically acceptable salts, solvates, hydrates, cage compounds, and polymorphs:

[0105]

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162] The stereocenter marked with an asterisk (*) is racemic or has (S)-stereochemistry, and compounds exhibiting (S)-stereochemistry may alternatively be racemic at that stereocenter.

[0163] In other embodiments, the compound of formula I is selected from the following compounds, their tautomers, pharmaceutically acceptable salts, solvates, hydrates, cage compounds, and polymorphs:

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217] The stereocenter marked with an asterisk (*) is racemic or has (S)-stereochemistry, and compounds exhibiting (S)-stereochemistry may alternatively be racemic at that stereocenter.

[0218] In some implementations, the stereocenter marked with an asterisk (*) has (S)-stereochemistry.

[0219] The compounds of Formula I are potent, selective NNMT inhibitors and are orally bioavailable.

[0220] In other embodiments, the triazolone NNMT inhibitor is selected from the following compounds, their tautomers, pharmaceutically acceptable salts, solvates, hydrates, cage compounds, and polymorphs:

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231] The stereocenter marked with an asterisk * is racemic or has (S)-stereochemistry, and compounds exhibiting (S)-stereochemistry may alternatively be racemic at the stereocenter, and Boc represents tert-butyloxycarbonyl.

[0232] Triazolone compounds of Formula I have a 1,3-disubstituted piperidinyl group. Alternatively, the 1,3-disubstituted piperidinyl group may be replaced by a 1,4-disubstituted piperidinyl group, a 1,3-disubstituted pyrrolidinyl group, or a 1,3-disubstituted azacyclic butyl group, and triazolone compounds having any of these three substituted groups are also disclosed herein.

[0233] Salt form of compounds

[0234] Compounds of Formula I have at least one basic nitrogen atom and can exist as a free base or as a salt. They can be used as a free base or as a pharmaceutically acceptable salt. The basic nitrogen atom can form an addition salt with an acid, such as an inorganic acid (e.g., HCl, HBr, HI, nitric acid, phosphoric acid, or sulfuric acid) or an organic acid (e.g., a carboxylic acid or a sulfonic acid). Acids suitable for preparing pharmaceutically acceptable salts include, but are not limited to, acetic acid, 2,2-dichloroacetic acid, acylated amino acids, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, boric acid, (+)-camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, cyclohexanesulfonic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactopyric acid, gentian acid, glucoheponic acid, D-gluconic acid, D-glucuronic acid, L-glutamic acid, α-oxoglutarate, and ethanol. Acids, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, (±)-DL-lactic acid, (+)-L-lactic acid, lactobionic acid, lauric acid, maleic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, perchloric acid, phosphoric acid, propionic acid, L-pyroglutamic acid, pyruvic acid, saccharic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (±)-DL-tartaric acid, (+)-L-tartaric acid, thiocyanate, p-toluenesulfonic acid, undecenoic acid, and valeric acid.

[0235] If a compound has an acidic group (e.g., a carboxyl group), that acidic group can form an addition salt with a base. Pharmaceutically acceptable base addition salts can be formed, for example, with a metal (e.g., an alkali metal or an alkaline earth metal) or an amine (e.g., an organic amine). Examples of metals that can be used as cations include, but are not limited to, alkali metals (e.g., lithium, sodium, potassium, and cesium), alkaline earth metals (e.g., magnesium, calcium, and barium), aluminum, and zinc. Metal cations can be provided, for example, by inorganic bases such as hydroxides, carbonates, and bicarbonates. Non-limiting examples of organic amines that can be used to form base addition salts include chloroprocaine, choline, cyclohexylamine, dibenzylamine, N,N'-dibenzylethylenediamine, dicyclohexylamine, diethanolamine, ethylenediamine, N-ethylpiperidine, histidine, isopropylamine, N-methylglucosamine, procaine, pyrazine, triethylamine, trimethylamine, and tromethamine. Pharmaceutically acceptable salts are discussed in detail in Handbook of Pharmaceutical Salts, Properties, Selection and Use, P. Stahl and C. Wermuth, Eds., Wiley-VCH (2011).

[0236] In some embodiments, the NNMT inhibitors described herein are used in the form of pharmaceutically acceptable salts. In some embodiments, the compounds are used in the form of hydrochloric acid (HCl) salts.

[0237] Pharmaceutical Composition

[0238] Additional embodiments of this disclosure relate to pharmaceutical compositions comprising the NNMT inhibitor described herein, or a tautomer thereof, a pharmaceutically acceptable salt, solvate, hydrate, cage compound, or polymorph, and one or more pharmaceutically acceptable excipients or carriers. The compositions may optionally contain additional therapeutic agents. The pharmaceutical composition contains a therapeutically effective amount or any suitable fraction thereof of an NNMT inhibitor, one or more pharmaceutically acceptable excipients or carriers, and optionally a therapeutically effective amount of additional therapeutic agent, and is formulated for administration to a subject for therapeutic use. With respect to the contents of the pharmaceutical composition, the terms “active ingredient,” “active agent,” “therapeutic agent,” “drug,” etc., encompass prodrugs.

[0239] The pharmaceutical composition contains an NNMT inhibitor in substantially pure form and an optional additional therapeutic agent. In some embodiments, the purity of the NNMT inhibitor and the optional additional therapeutic agent is independently at least about 95%, about 96%, about 97%, about 98%, or about 99%. Furthermore, the pharmaceutical composition is substantially free of contaminants or impurities. In some embodiments, the level of contaminants or impurities in the pharmaceutical composition, excluding residual solvents, does not exceed about 5%, about 4%, about 3%, about 2%, or about 1% of the combined weight of the intended active ingredient and the inactive ingredient. The pharmaceutical composition is generally prepared according to current Good Manufacturing Practices (GMP), such as those recommended or required by, for example, the Federal Food, Drug, and Cosmetic Act §501(a)(2)(B) and the International Conference on Harmonisation Q7 guidance.

[0240] Pharmaceutical compositions / formulations can be prepared in a sterile form. For example, pharmaceutical compositions / formulations intended for parenteral administration by injection or infusion are typically sterile. Sterile pharmaceutical compositions / formulations are formulated or manufactured according to pharmaceutical-grade sterilization standards known to those skilled in the art, such as those disclosed or required in United States Pharmacopeia Chapters 797, 1072, and 1211 and Title 21, Section 211 of the Federal Regulations.

[0241] Pharmaceutically acceptable excipients and carriers include pharmaceutically acceptable substances, materials, and media. Non-limiting examples of excipient types include liquid and solid fillers, diluents, binders, lubricants, flow aids, surfactants, dispersants, disintegrants, emulsifiers, wetting agents, suspending agents, thickeners, solvents, isotonic agents, buffers, pH adjusters, absorption delay agents, stabilizers, antioxidants, preservatives, antimicrobial agents, antibacterial agents, antifungal agents, chelating agents, adjuvants, sweeteners, flavoring agents, colorants, encapsulation materials, and coating materials. The use of such excipients in pharmaceutical formulations is known in the art. For example, conventional media and carriers include, but are not limited to, oils (e.g., vegetable oils such as olive oil and sesame oil), aqueous solvents {e.g., saline, buffered saline (e.g., phosphate-buffered saline [PBS]), and isotonic solutions (e.g., Ringer's solution)}, and organic solvents (e.g., dimethyl sulfoxide [DMSO] and alcohols [e.g., ethanol, glycerol, and propylene glycol]). This disclosure covers the use of conventional excipients and carriers in formulations containing NNMT inhibitors unless any conventional excipient or carrier is incompatible with the active ingredient. See, for example, Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins (Philadelphia, Pennsylvania) (2005); Handbook of Pharmaceutical Excipients, 5th Ed., Rowe et al., Eds., The Pharmaceutical Press and the American Pharmaceutical Association (2005); Handbook of Pharmaceutical Additives, 3rd Ed., Ash and Ash, Eds., Gower Publishing Co. (2007); and Pharmaceutical Pre-formulation and Formulation, Gibson, Ed., CRC Press (Boca Raton, Florida) (2004).

[0242] Appropriate formulations may depend on various factors, such as the chosen route of administration. Potential routes of administration for pharmaceutical compositions containing NNMT inhibitors include, but are not limited to, oral, parenteral (including intradermal, subcutaneous, intramuscular, intravascular, intravenous, intraarterial, peritoneal, intracavitary, intramedullary, intrathecal, and local), and local (including dermal / epidermal, transdermal, mucosal, transmucosal, intranasal [e.g., via nasal spray or drops], ocular / intraocular [e.g., via eye drops], pulmonary [e.g., via oral or nasal inhalation], buccal, sublingual, rectal [e.g., via suppository], and vaginal [e.g., via suppository]). Local formulations may be designed to produce local or systemic therapeutic effects. In some embodiments, NNMT inhibitors are administered in oral dosage forms such as tablets, capsules, or pills.

[0243] For example, NNMT inhibitor formulations suitable for oral administration may be presented in discrete unit form suitable for immediate, controlled, or sustained release, such as pellets, capsules (including push-in capsules and soft capsules), tablets, pills, cachets, or lozenges; as powders or granules; as semi-solids, sugar baits, pastes, or gels; as solutions or suspensions in aqueous and / or non-aqueous liquids; or as oil-in-water or water-in-oil liquid emulsions.

[0244] Push-in capsules or two-piece hard gelatin capsules may contain an NNMT inhibitor mixed with, for example, a filler or inert solid diluent (e.g., calcium carbonate, calcium phosphate, kaolin, or lactose), a binder (e.g., starch), a flow aid or lubricant (e.g., talc or magnesium stearate), and a disintegrant (e.g., cropovidone), and optionally a stabilizer and / or preservative. For soft capsules or single-piece gelatin capsules, the NNMT inhibitor may be dissolved or suspended in a suitable liquid (e.g., liquid polyethylene glycol or an oily medium such as fatty oil, peanut oil, olive oil, or liquid paraffin), and liquid-filled capsules may contain one or more other liquid excipients and / or semi-solid excipients, such as stabilizers and / or amphiphilic agents (e.g., fatty acid esters of glycerol, propylene glycol, or sorbitol).

[0245] Tablets may contain NNMT inhibitors, which are mixed with, for example, fillers or inert diluents (e.g., calcium carbonate, calcium phosphate, lactose, mannitol, or microcrystalline cellulose), binders (e.g., starch, gelatin, gum arabic, alginate or its salts, or microcrystalline cellulose), lubricants (e.g., stearic acid, magnesium stearate, talc, or silica), and disintegrants (e.g., crospovidone, croscarmellose sodium, or colloidal silica), and optionally contain surfactants (e.g., sodium dodecyl sulfate). Tablets may be uncoated, or may be coated with, for example, enteric coatings that protect the active agent from the acidic environment of the stomach, or with materials that delay the disintegration and absorption of the active agent in the gastrointestinal (GI) tract, thereby providing sustained action over a longer period of time.

[0246] Compositions for oral administration may also be formulated as solutions or suspensions in aqueous and / or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions. Dispersible powders or particles of NNMT inhibitors may be mixed with aqueous liquids, organic solvents and / or oils, and any suitable excipients (e.g., any combination of dispersants, wetting agents, suspending agents, emulsifiers, and / or preservatives) to form solutions, suspensions, or emulsions.

[0247] NNMT inhibitors can also be formulated for parenteral administration by injection or infusion to bypass gastrointestinal absorption and first-pass metabolism. An exemplary parenteral route is intravenous administration. Additional advantages of intravenous administration include direct administration of the therapeutic agent into the systemic circulation for rapid systemic effects, and the ability to administer the agent continuously or / and in large volumes if needed. Formulations for injection or infusion can be in the form of solutions, suspensions, or emulsions, for example, in oily or aqueous media, and may contain excipients such as suspending agents, dispersants, and / or stabilizers. For example, aqueous or non-aqueous (e.g., oily) sterile injectable solutions may contain NNMT inhibitors as well as excipients such as antioxidants, buffers, antibacterial agents, and solutes that make the formulation isotonic with the patient's blood. Aqueous or non-aqueous sterile suspensions may contain NNMT inhibitors as well as excipients such as suspending agents and thickeners, and optionally contain stabilizers and reagents that increase the solubility of the NNMT inhibitor to allow for the preparation of more concentrated solutions or suspensions. Another example is that sterile aqueous solutions for injection or infusion (e.g., subcutaneous or intravenous) may contain NNMT inhibitors, isotonic agents (e.g., sodium chloride), buffers (e.g., sodium citrate), preservatives (e.g., m-cresol), and optionally bases (e.g., NaOH) and / or acids (e.g., HCl) to adjust the pH.

[0248] Topical formulations for application to the skin or mucous membranes can deliver therapeutic agents locally to the site of action or into the bloodstream for systemic distribution. Advantages of topical application may include bypassing the gastrointestinal tract (including enzymes and acids in the gastrointestinal tract and absorption therethrough) and first-pass metabolism; delivery of therapeutic agents with short half-lives, low therapeutic indices, and / or low oral bioavailability; controlled, continuous, and sustained release of the therapeutic agent; more uniform plasma levels or delivery profiles; lower doses and lower frequencies of administration; reduced systemic side effects (e.g., side effects caused by transient overdose or excessively high peak plasma drug concentrations); minimally invasive or non-invasive nature; ease of self-administration; and improved patient compliance.

[0249] Compositions suitable for topical application include, but are not limited to, liquid or semi-liquid formulations such as sprays, gels, liniments, lotions, oil-in-water or water-in-oil emulsions such as creams, foams, ointments, and pastes, and solutions or suspensions such as drops (e.g., eye drops, nasal drops, and ear drops). See Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins (Philadelphia, Pennsylvania

[2005] ). Various excipients may be included in topical formulations. For example, solvents, including appropriate amounts of alcohol, may be used to dissolve active agents. Other optional excipients include, but are not limited to, gelling agents, thickeners, emulsifiers, surfactants, stabilizers, buffers, antioxidants, preservatives, coolants (e.g., menthol), opacifiers, fragrances, and colorants. For active agents with low skin or mucous membrane penetration rates, topical formulations may contain chemical penetration enhancers (e.g., fatty acid esters [e.g., isopropyl myristate or isopropyl palmitate], fatty acids [e.g., palmitic acid, oleic acid, or palmitoleic acid], and / or alcohols [e.g., propylene glycol or fatty alcohols such as geraniol or farnesol]) to increase the penetration of the active agent through the skin or mucous membranes. Topical formulations may also contain irritation-reducing excipients that reduce any irritation to the skin or mucous membranes caused by the active agent, chemical penetration enhancer, or any other component of the formulation. In some embodiments, the topical composition comprises a therapeutic agent dissolved, dispersed, or suspended in a carrier. The carrier may be in the form of, for example, a solution, suspension, emulsion, ointment, or gel matrix, and may contain, for example, petrolatum, lanolin, waxes (e.g., beeswax), mineral oil, long-chain alcohols, polyethylene glycol or polypropylene glycol, diluents (e.g., water and / or alcohols [e.g., ethanol or propylene glycol]), gels, emulsifiers, thickeners, stabilizers, or preservatives, or any combination thereof. Topical formulations can be administered via transdermal or transmucosal delivery devices such as transdermal patches, microneedle patches, or iontophoresis devices. Topical compositions can be delivered transdermally or transmucosally via concentration gradients (with or without chemical penetration enhancers) or active mechanisms (e.g., iontophoresis or microneedles).

[0250] For topical application, NNMT inhibitors can be formulated as, for example, buccal or sublingual tablets or pills. The advantages of buccal or sublingual tablets or pills include avoiding gastrointestinal absorption and first-pass metabolism, as well as rapid absorption into the systemic circulation. Buccal or sublingual tablets or pills can be designed to provide a faster release of the NNMT inhibitor for more rapid uptake into the systemic circulation. In addition to a therapeutically effective amount of the NNMT inhibitor, buccal or sublingual tablets or pills may contain any combination of suitable excipients, including but not limited to fillers and diluents (e.g., mannitol and sorbitol), binders (e.g., sodium carbonate), wetting agents (e.g., sodium carbonate), disintegrants (e.g., crospovidone and croscarmellose sodium), lubricants (e.g., silica [including colloidal silica] and sodium stearate fumarate), stabilizers (e.g., sodium bicarbonate), flavoring agents (e.g., spearmint flavor), sweeteners (e.g., sucralose), and coloring agents (e.g., iron oxide yellow).

[0251] For topical application, NNMT inhibitors can also be formulated for intranasal administration. The nasal mucosa provides a large surface area, porous endothelium, a highly vascularized subepithelial layer, and high absorption, thus resulting in high bioavailability. Furthermore, intranasal administration avoids first-pass metabolism and allows for the introduction of significant concentrations of NNMT inhibitors into the central nervous system (CNS). Intranasal formulations may contain NNMT inhibitors along with excipients such as solubility enhancers (e.g., propylene glycol), humectants (e.g., mannitol or sorbitol), buffers, and water, and optionally preservatives (e.g., benzalkonium chloride), mucosal adhesives (e.g., hydroxyethyl cellulose), and / or penetration enhancers. Intranasal solutions or suspensions may be administered into the nasal cavity by any suitable means, including but not limited to droppers, pipettes, or sprays using, for example, metered nebulizer pumps. Table 1 shows exemplary excipients for nasal spray formulations.

[0252] Another method of local administration of NNMT inhibitors is pulmonary administration, including oral and nasal inhalation. Pulmonary-administered drugs can treat lung diseases and / or systemic diseases because the lungs act as a gateway to systemic circulation. Advantages of pulmonary drug delivery include, for example: 1) avoidance of first-pass metabolism; 2) rapid drug action; 3) large absorption surface area in the alveolar region, high lung permeability (thin air-blood barrier), and rich airway vascularity; 4) reduced extracellular enzyme levels due to the large alveolar surface area compared to the gastrointestinal tract; and 5) smaller doses required to achieve equivalent therapeutic effects compared to other oral routes, resulting in fewer systemic side effects. Oral inhalation also allows the drug to act more rapidly in the CNS. One advantage of oral inhalation over nasal inhalation includes deeper penetration / deposition of the drug into the lungs, although nasal inhalation can also deliver the drug into systemic circulation through the mucous membranes of the nasal cavity and lungs. Oral or nasal inhalation can be achieved, for example, by metered-dose inhalers (MDIs), dry powder inhalers (DPIs), or nebulizers, as known in the art. In some embodiments, the sterile aqueous solution for oral inhalation contains an NNMT inhibitor, sodium chloride, a buffer (e.g., sodium citrate), optionally a preservative (e.g., m-cresol), and optionally a base (e.g., NaOH) and / or an acid (e.g., HCl) to adjust the pH. Table 1 shows other exemplary excipients for oral inhalation formulations.

[0253] Table 1. Exemplary excipients and carriers for pulmonary and nasal formulations

[0254]

[0255] In some embodiments, NNMT inhibitors are administered transdermally. In some embodiments, the topical composition or transdermal delivery system comprises a chemical penetration enhancer (e.g., a surfactant [e.g., sodium lauryl ether sulfate], optionally in combination with an aromatic compound [e.g., phenylpiperazine]) that facilitates the transport of the NNMT inhibitor across the skin. In further embodiments, the NNMT inhibitor is administered via a transdermal patch. In some embodiments, the transdermal patch is a reservoir-type patch comprising an impermeable backing layer / membrane, a liquid- or gel-based drug reservoir, a semipermeable membrane acting as a rate-limiting or controlled-release diffusion barrier, and a skin-contact adhesive layer. The semipermeable membrane may be composed of, for example, suitable polymeric materials such as nitrocellulose or cellulose acetate, polyisobutylene, polypropylene, polyvinyl acetate, or polycarbonate. In other embodiments, the transdermal patch is a patch of drug in an adhesive comprising an impermeable backing layer / membrane and a skin-contact adhesive layer incorporating the drug into a polymer or adhesive binder. The adhesive for the drug-loaded skin contact adhesive layer can be, for example, a pressure-sensitive adhesive (PSA), such as a PSA composed of an acrylic polymer (e.g., polyacrylate), a polyalkylene polymer (e.g., polyisobutylene), or a silicone-based polymer (e.g., silicone-2675 or silicone-2920). Transdermal drug delivery systems, including patches, can be designed to provide controlled and prolonged drug release over a period of about 1 week, about 2 weeks, about 3 weeks, about 1 month, or longer than 1 month.

[0256] In some embodiments, the NNMT inhibitor is delivered from a sustained-release composition. As used herein, the term "sustained-release composition" encompasses sustained-release, extended-release, prolonged-release, delayed-release, sustained-release, and controlled-release compositions, systems, and devices. The advantages of sustained-release compositions include, but are not limited to, more uniform blood drug levels (e.g., avoiding wide peak-to-trough fluctuations), delivery of a therapeutically effective amount of drug over an extended period, reduced dosing frequency, and reduced side effects (e.g., avoiding drug overdose). In some embodiments, the sustained-release composition delivers the NNMT inhibitor over a period of at least about 1 day, about 2 days, about 3 days, about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, about 3 months, or longer than 3 months.

[0257] In some embodiments, the sustained-release composition is a drug encapsulation system, such as nanoparticles, microparticles, or capsules made of, for example, biodegradable polymers and / or hydrogels. In some embodiments, the sustained-release composition comprises a hydrogel. Non-limiting examples of polymers that can be composed of hydrogels include polyvinyl alcohol, acrylate polymers (e.g., sodium polyacrylate), and other homopolymers and copolymers having a relatively large number of hydrophilic groups (e.g., hydroxyl and / or carboxylate groups). In other embodiments, the sustained-release drug encapsulation system comprises a membrane-closed reservoir containing a drug and the membrane is permeable to the drug. Such a drug delivery system can be, for example, in the form of a transdermal patch.

[0258] In some embodiments, the sustained-release composition is formulated as polymer nanoparticles or microparticles, wherein the polymer particles can be delivered, for example, by injection or from an implant. In some embodiments, the polymer implant or polymer nanoparticles or microparticles are composed of a biodegradable polymer. In some embodiments, the biodegradable polymer comprises lactic acid and / or glycolic acid [e.g., L-lactic acid-based copolymers, such as poly(L-lactide-co-glycolic acid) or poly(L-lactic acid-co-D,L-2-hydroxyoctanoic acid)]. For example, biodegradable polymer microspheres composed of polylactic acid and / or polyglycolic acid can be used as a sustained-release pulmonary drug delivery system. The biodegradable polymer of the polymer implant or polymer nanoparticles or microparticles can be selected such that the polymer is substantially completely degraded around the expected end of the treatment period, and such that the byproducts of polymer degradation are biocompatible as the polymer itself.

[0259] In further embodiments, the sustained-release composition comprises a dendritic polymer. In some embodiments, the dendritic polymer is a water-soluble dendritic polymer, such as a poly(amidoamine) (PAMAM) dendritic polymer. In some embodiments, the dendritic polymer encapsulates the drug by forming a dendritic polymer-drug supramolecular assembly. In other embodiments, the sustained-release composition comprises a water-soluble polymer [e.g., poly(DL-lactide)] or liposomes encapsulating a drug complexed with a dendritic polymer.

[0260] In other embodiments, the sustained-release composition is an oral dosage form, such as a tablet or capsule. For example, the drug may be encapsulated in an insoluble porous matrix such that the dissolved drug must exit the matrix before it can be absorbed through the gastrointestinal tract. Alternatively, the drug may be encapsulated in a matrix that swells to form a gel, through which the drug flows out. Sustained release can also be achieved using a single-layer or multi-layer osmotic controlled-release oral delivery system (OROS). An OROS is a tablet having a semi-permeable outer membrane and one or more laser-drilled pores therein. As the tablet passes through the body, water is absorbed through the semi-permeable membrane via osmosis, and the resulting osmotic pressure pushes the drug through the pores in the tablet into the gastrointestinal tract, where it is absorbed.

[0261] For delayed or sustained release of NNMT inhibitors, the composition can also be formulated as a reservoir, for example, that can be implanted or injected into the body, such as intramuscular, intradermal, or subcutaneous injection. The reservoir formulation can be designed to deliver the NNMT inhibitor over an extended period of time, such as at least about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 6 weeks, about 2 months, about 3 months, or longer than 3 months. For example, the NNMT inhibitor can be formulated with polymeric materials (e.g., polyethylene glycol [PEG], polylactic acid [PLA], or polyglycolic acid [PGA], or copolymers thereof [e.g., PLGA or PLA-PEG]), hydrophobic materials (e.g., as an emulsion in oil), and / or ion exchange resins, as more lipophilic derivatives (e.g., as a derivative with fatty acids such as C8-C). 20 Esters or salts of fatty acids (e.g., decanoic acid), or as slightly soluble derivatives (e.g., slightly soluble salts). As an illustrative example, NNMT inhibitors can be incorporated or encapsulated in sustained-release microparticles composed of PLGA and formulated as a once-monthly reservoir.

[0262] NNMT inhibitors can also be contained or dispersed in a matrix material. The matrix material may contain a polymer (e.g., ethylene-vinyl acetate) and control drug release by controlling the dissolution and / or diffusion of the drug from, for example, a reservoir, and may enhance the stability of the drug when contained within the reservoir. Such a release system can be designed as a sustained-release system, configured as, for example, a transdermal or transmucosal patch, and may contain excipients that accelerate drug release, such as water-swellable materials (e.g., hydrogels) that facilitate the removal of the drug from the reservoir. Examples of such release systems are described in U.S. Patents 4,144,317 and 5,797,898.

[0263] When a time-varying variation in plasma levels is required, the release system can provide a time-modulated release profile (e.g., pulsed release), or a more continuous or consistent release profile when a constant plasma level is required. Pulsed release can be achieved from a single reservoir or multiple reservoirs. For example, in the case where each reservoir provides a single pulse, multiple pulses can be achieved by releasing single pulses from each of the multiple reservoirs in a time-staggered manner (“pulsed” release). Alternatively, multiple pulses from a single reservoir can be achieved by incorporating multiple layers of the release system and other materials into a single reservoir. Continuous release can be achieved by incorporating a release system that degrades, dissolves, or allows the drug to diffuse over an extended time. Furthermore, continuous release can be approximated by rapidly and continuously releasing multiple drug pulses (“digital” release). Active release systems can be used alone or in combination with passive release systems, as described in U.S. Patent 5,797,898.

[0264] Furthermore, pharmaceutical compositions containing NNMT inhibitors can be formulated as, for example, liposomes, micelles (e.g., those composed of biodegradable natural and / or synthetic polymers such as lactosomes), nanoparticles (e.g., lipid nanoparticles such as solid lipid nanoparticles), microparticles, or microspheres, whether or not they are designed for sustained release. For example, liposomes can be used as sustained-release pulmonary drug delivery systems to deliver drugs to the alveolar surface for the treatment of lung and systemic diseases. In another example, lipid nanoparticles containing lipophilic drugs can be delivered to the lungs via oral inhalation for the treatment of lung or systemic diseases.

[0265] In some embodiments, the liposomes or micelles are composed of one or more phospholipids. Phospholipids include, but are not limited to, phosphatidic acids (e.g., DEPA, DLPA, DMPA, DOPA, DPPA, and DSPA), phosphatidylcholine (e.g., DDPC, DEPC, DLPC, DLOPC, DMPC, DOPC, DPPC, DSPC, MPPC, MSPC, PLPC, PMPC, POPC, PSPC, SMPC, SOPC, and SPPC), phosphatidylethanolamine (e.g., DEPE, DLPE, DMPE, DOPE, DPPE, DSPE, and POPE), phosphatidylglycerol (e.g., DEPG, DLPG, DMPG, DOPG, DPPG, DSPG, and POPG), phosphatidylserine (e.g., DLPS, DMPS, DOPS, DPPS, and DSPS) and their salts (e.g., sodium and ammonium salts). In some embodiments, the liposomes or micelles are composed of one or more phosphatidylcholine. Liposomes have a hydrophilic core, making them particularly suitable for delivering more hydrophilic drugs, while micelles have a hydrophobic core, making them particularly suitable for delivering more hydrophobic drugs. Both liposomes and micelles can permeate across biological membranes. Furthermore, liposomes and micelles composed of fused lipids (e.g., DPPG) can fuse with the cell membrane, thereby delivering drugs into those cells. Sustained drug release can be provided by liposomes and micelles, in part, based on their degradation rates.

[0266] Pharmaceutical compositions can be prepared in any suitable manner known in the art, such as by conventional methods of mixing, dissolving, suspending, granulating, making sugar-coated pellets, grinding, emulsifying, encapsulating, embedding or pressing, or any combination thereof.

[0267] Compositions can be presented in unit dosage forms, in which all active and inactive ingredients are combined in a suitable system and the composition to be administered can be formed without mixing the components. Unit dosage forms typically contain a therapeutically effective dose of the drug, but may contain appropriate fractions such that multiple unit dosage forms are taken to achieve a therapeutically effective dose. Examples of unit dosage forms include tablets, capsules, or pills for oral administration; solutions in pre-filled syringes of single-use pens or pens with dose counters for parenteral (e.g., intravenous, subcutaneous, or intramuscular) injection; capsules, cartridges, or blister packs pre-filled in inhalers or manually loaded into inhalers; and reservoir-type transdermal patches or patches containing the drug in an adhesive.

[0268] Alternatively, the composition may be presented as a kit in which the active ingredient, excipients, and carrier [e.g., solvent] are provided in two or more separate containers (e.g., ampoules, vials, tubes, bottles, or syringes) and need to be combined to form the composition to be administered. The kit may include instructions for storing, preparing, and administering the composition (e.g., a solution for parenteral injection).

[0269] The kit may contain all active and inactive ingredients in a single dosage form, or active and inactive ingredients in two or more separate containers, and may include instructions for use of the pharmaceutical composition to treat a medical condition. The kit may further include a device for delivering the composition, such as a needle and syringe, injection pen, inhaler, or transdermal patch.

[0270] In some embodiments, the kit contains an NNMT inhibitor or a pharmaceutically acceptable salt, solvate, hydrate, cage, or polymorph thereof, or a pharmaceutical composition comprising the thereof, along with instructions for use of the NNMT inhibitor or composition to treat a medical condition (e.g., tumor or cancer, metabolic disorder, or liver disease). In some embodiments, the kit also includes a device for delivering the NNMT inhibitor or composition, such as an injection pen, inhaler, or transdermal patch.

[0271] Therapeutic uses of NNMT inhibitors

[0272] High expression of nicotinamide N-methyltransferase (NNMT) is a predictor of poor prognosis in many types of human cancer [Cao et al., Frontiers in Genetics, 13:1000515 (2022); and Dang et al., Open Medicine, 17:292-303 (2022)]. Elevated NNMT levels or activity promote cancer cell survival, growth, proliferation, migration, invasion and metastasis, as well as resistance to chemotherapeutic drugs [Li et al., Frontiers in Oncology, 12:894744 (2022); Pozzi et al., Biomolecules, 12:1173 (2022); and Eckert et al., Nature, 569:723-728 (2019)].

[0273] In some embodiments, the NNMT inhibitors disclosed herein are used to treat tumors or cancers. In some embodiments, the tumor or cancer is selected from solid tumors, head and neck tumors and cancers (e.g., oral squamous cell carcinoma, esophageal squamous cell carcinoma, nasopharyngeal carcinoma, and papillary thyroid carcinoma), CNS tumors and cancers (e.g., glioblastoma, glioma, and neuroblastoma), lung cancer (e.g., non-small cell lung cancer), gynecological malignancies (e.g., breast cancer, ovarian cancer [e.g., high-grade serous carcinoma], and uterine cancer [e.g., cervical cancer and endometrial cancer]), and gastrointestinal cancers (e.g., stomach cancer [e.g., malignant epithelial tumors of the stomach]). Cancers include carcinoma (and colorectal cancer), liver cancer (e.g., hepatocellular carcinoma and intrahepatic cholangiocarcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), urinary system cancers (e.g., kidney cancer [e.g., clear cell renal cell carcinoma and papillary renal cell carcinoma], bladder cancer and prostate cancer), skin cancer (e.g., melanoma), sarcoma, and blood and lymphomas (e.g., leukemia and lymphoma [e.g., non-Hodgkin lymphoma such as diffuse large B-cell lymphoma]). In some embodiments, the tumor or cancer is characterized by intraperitoneal, peritoneal, and / or omental metastases, such as ovarian cancer (e.g., epithelial ovarian cancer), endometrial cancer, gastrointestinal cancer (e.g., gastric cancer or colorectal cancer), or pancreatic cancer (e.g., pancreatic ductal adenocarcinoma).

[0274] Furthermore, overexpression of NNMT in human adipose tissue is associated with obesity and insulin resistance [Pissios, Trends Endocrinol. Metab., 28(5):340-353 (2017)]. Therefore, in further embodiments, the NNMT inhibitors disclosed herein are used to treat metabolic disorders. In some embodiments, the metabolic disorder is selected from obesity, obesity-related disorders (e.g., cardiovascular disorders and hypertension), hyperlipidemia (e.g., hypercholesterolemia and hypertriglyceridemia), metabolic syndrome, hyperglycemia, impaired glucose tolerance, insulin resistance, type 2 diabetes, polycystic ovary syndrome (PCOS), and age-related disorders (e.g., sarcopenia). In some embodiments, the metabolic disorder is obesity, obesity-related disorders, or type 2 diabetes.

[0275] Obesity is a major risk factor for a variety of disorders. Obesity-related disorders include, but are not limited to, insulin resistance, type 2 diabetes, cardiovascular diseases (e.g., coronary artery disease, atherosclerosis, peripheral vascular disease, hyperlipidemia, and hypertension), cerebrovascular diseases (e.g., stroke), inflammatory diseases (e.g., arthritis such as osteoarthritis), fatty liver diseases (e.g., NAFLD and NASH), obesity-related cancers (e.g., breast cancer, colorectal cancer, kidney cancer, and prostate cancer), PCOS, and sleep apnea.

[0276] Age-related disorders and conditions include, but are not limited to, aging, hypertension, eye diseases (e.g., age-related macular degeneration [AMD], cataracts, and keratoconjunctivitis sicca [dry eye]), hearing loss, bone diseases (e.g., osteoporosis), muscle diseases (e.g., muscle atrophy and sarcopenia), neurodegenerative diseases (e.g., dementia [e.g., Alzheimer's disease] and Parkinson's disease), metabolic disorders (e.g., metabolic degeneration, metabolic syndrome, diabetes [including type 1 and type 2 diabetes] and obesity), cardiovascular diseases (e.g., arteriosclerosis), inflammatory diseases (e.g., chronic inflammation, arthritis, and chronic obstructive pulmonary disease [COPD]), fibrotic diseases (e.g., idiopathic pulmonary fibrosis [IPF]), DNA repair syndromes (e.g., Cockayne syndrome), and tumors and cancers.

[0277] Furthermore, NNMT is upregulated in hepatic steatosis, hepatitis, liver fibrosis, and cirrhosis [Liang et al., Genes & Diseases, 10:1883-1893 (2023)]. Therefore, in additional embodiments, the NNMT inhibitors disclosed herein are used to treat liver diseases. In some embodiments, the liver disease is selected from hepatic steatosis, alcoholic liver disease (ALD), non-alcoholic fatty liver disease (NAFLD), hepatitis, alcoholic steatohepatitis (ASH), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis. In some embodiments, the liver disease is ALD, NAFLD, ASH, or NASH.

[0278] The therapeutically effective dose and frequency of administration of NNMT inhibitors can depend on a variety of factors, including the medical condition being treated, the severity of the condition, the potency of the compound, the route of administration, the subject's age, weight, general health condition, sex and diet, and the subject's response to treatment, and can be determined by the attending physician. In some embodiments, the effective daily dose (e.g., maintenance dose) of an NNMT inhibitor is about 1 mg to about 200 mg, about 1 mg to about 50 mg, about 50 mg to about 100 mg, about 100 mg to about 150 mg, or about 150 mg to about 200 mg, or as determined by the attending physician, and can be administered as a single dose or in divided doses (e.g., twice, three, or four times daily). In some embodiments, the effective daily dose (e.g., maintenance dose) of an NNMT inhibitor is about 1 mg to about 25 mg or about 25 mg to about 50 mg. The frequency of administration can depend on, for example, the chosen route of administration. For example, administration via the lungs (e.g., via oral inhalation) may be more frequent (e.g., twice, three, or four times daily). To establish therapeutic levels of NNMT inhibitors more quickly, an initial loading dose of the inhibitor (e.g., about 2 or 3 times the maintenance dose) larger than the maintenance dose can be administered at the start of treatment (e.g., during the first three days of treatment), followed by the maintenance dose.

[0279] NNMT inhibitors can be administered via any suitable route, which may depend on factors such as the medical condition being treated and its location, as well as the pharmacokinetics of the inhibitor. Potential routes of administration for NNMT inhibitors include, but are not limited to, oral, parenteral (including intradermal, subcutaneous, intramuscular, intravascular, intravenous, intra-arterial, peritoneal, intracavitary, intramedullary, intrathecal, and local), and local (including dermal / epidermal, transdermal, mucosal, transmucosal, intranasal [e.g., via nasal spray or drops], ocular / intraocular [e.g., via eye drops], pulmonary [e.g., via oral or nasal inhalation], buccal, sublingual, rectal [e.g., via suppository], and vaginal [e.g., via suppository]).

[0280] In some embodiments, the NNMT inhibitor is administered orally. In other embodiments, the NNMT inhibitor is administered parenterally, such as intravenously, subcutaneously, or intramuscularly. In a further embodiment, the NNMT inhibitor is administered intratumorally to treat tumors or cancer.

[0281] The duration of treatment with an NNMT inhibitor can be determined by the attending physician to achieve the desired outcome. In some implementations, the NNMT inhibitor is administered for at least about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks (1 month). In other implementations, the NNMT inhibitor is administered for at least about 6 weeks, about 2 months, about 3 months, about 6 months, about 1 year, about 2 years, about 3 years, or longer than 3 years.

[0282] In some embodiments, it is disclosed to modulate NAD using an NNMT inhibitor having Formula I or a tautomer thereof or a pharmaceutically acceptable salt thereof. + Methods for metabolites in the methionine and S-adenosine-L-methionine (SAM) / S-adenosine-L-homocysteine ​​(SAH) pathways.

[0283] In some implementations, methods for regulating gene expression or epigenomes are disclosed using an NNMT inhibitor having Formula I or a tautomer thereof or a pharmaceutically acceptable salt thereof.

[0284] combination therapy

[0285] The NNMT inhibitors disclosed herein can optionally be used in combination with one or more additional therapeutic agents to treat medical conditions. In some embodiments, the NNMT inhibitors are used in combination with one or more additional antitumor / anticancer agents to treat tumors or cancers.

[0286] Antitumor / anticancer agents include, but are not limited to:

[0287] 1) Cytotoxic agents, including but not limited to:

[0288] i) Alkylating agents, such as aziridines (e.g., diazinon, mitomycin, and thiotepa), nitrogen mustards (e.g., mannomustine, nitrogen mustard (dichloromethyldiethylamine or bis(2-chloroethyl)methylamine), aniline mustard, bendamustine, benzoic acid mustard, chlorinated nitrogen mustard, C6-galactosamine, melphalan, ossichlorin (nitromin), prednimustine, uracil nitrogen mustard, carbamate nitrogen mustards [e.g., estradiol], and oxazaphosphorines [e.g., cyclophosphamide]. Amines, ifosfamide, maphosphamide and trefophosphatamide), nitrosoureas (e.g., carmustine, formustine, lomustine, nimustine, N-nitroso-N-methylurea, ramustine, semustine and streptozotocin), platinum-containing compounds (e.g., cisplatin, carboplatin and oxaliplatin), alkyl sulfonates (e.g., busulfan, mannosulfan and sulfan), hydrazides (e.g., dacarbazine and procarbazine), imidazotetraazines (e.g., mitoxazole and temozazole) and triazines (e.g., hexamethylmelamine [hexamethylcyanamide]);

[0289] ii) Cytotoxic antibiotics, such as anthracyclines (e.g., arubicin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, pirarubicin and pentorubicin), actinomycins (e.g., actinomycin D), bleomycins (e.g., bleomycin A2 and B2), mitomycins (e.g., mitomycin C) and procainamides;

[0290] iii) Antimetabolites, such as antifolate agents (e.g., aminopterin, methotrexate, pemetrexed, and pralatrexate), deoxynucleoside analogs (e.g., 5-azacytidine [azacytidine], 5-aza-2'-deoxycytidine [decitabine], cladribine, clofarabine, cytarabine, decitabine, fludarabine, gemcitabine, nelabine, and pentostatin), fluoropyrimidines (e.g., 5-fluorouracil, 1-hexylcarbamoyl-5-fluorouracil [camofluorine], capecitabine, 5-fluoro-5'-deoxyuridine [doxyfluuridine], and trifluorouridine), and thiopurines (e.g., thioguanine, azathiopurine, and 6-mercaptopurine);

[0291] iv) Anti-microtubule agents, such as salipodoxins (e.g., salipodoxins 15), epokines (e.g., epokine AF), sponginoids (e.g., sponginoid B) and their analogues (e.g., iribulin), maytansin, maytansinoids (e.g., emtansine, maytansin, ravtansine and soravtansine), taxanes (e.g., paclitaxel, docetaxel and carbamazepine), vinca alkaloids (e.g., vincaine, vincristine, vindesin, vinflunine and vinorelbine), colchicine, nocodazole, podophyllotoxin and rhizomycin;

[0292] v) Histone deacetylase inhibitors, such as trichostatins (e.g., trichostatin A), romidesin, pabistat, and vorinostat;

[0293] vi) Kinase inhibitors, such as bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, vemodil, curcumin, cyclocreatine, rotenone, foctocrisin, milkweed alkaloids, astrocytocin and its derivatives (e.g., midostaurin) and tyrosine phosphorylation inhibitors (e.g., tyrosine phosphorylation inhibitors AG 34 and AG 879);

[0294] vii) Topoisomerase I inhibitors, such as camptothecin, irinotecan and topotecan;

[0295] viii) Topoisomerase II targets, such as topoisomerase II poisons (e.g., etoposide, tafluposide, teniposide, doxorubicin and mitoxantrone) and topoisomerase II inhibitors (e.g. neomycin, meparone and azorubicin).

[0296] ix) DNA or RNA synthesis inhibitors, such as 3-amino-1,2,4-benzotriazine 1,4-dioxide, cytosine β-D-arabinofuranyl glucoside, 5,6-dichlorobenzimidazole 1-β-D-furanoside, ganciclovir and hydroxyurea;

[0297] x) Protein synthesis inhibitors, such as homoharringtonine;

[0298] xi) Cell growth and differentiation regulators, such as retinoids (e.g., all-trans retinol [vitamin A], 11-cis retinol, all-trans retinal [vitamin A aldehyde], 11-cis retinal, all-trans retinoic acid [retinoic acid], 9-cis-retinoic acid [alvitamin A acid], 11-cis-retinoic acid, 13-cis-retinoic acid [isotretinoin], all-trans retinyl ester, ectretinoic acid ester, acitretin, adapalene, bexarotine, and tazarotene);

[0299] xii) Cell proliferation inhibitors, such as mTOR inhibitors (e.g., everolimus, nomulix, lidafomus, sirolimus [rapamycin], tesirobolimus, umimilimus [biolimus A9] and zotalimus), apigenin, cholecalciferol (vitamin D3) and sex hormone-binding globulin;

[0300] xiii) Apoptosis inducers, such as 17-allylamino-17-demethoxygerdin, melatonin, mevinoxin, psoralen, carotenoids, troglitazone, histone deacetylase inhibitors (e.g., romidisin), and retinoid X receptor (RXR) agonists (e.g., retinoids such as bexarotine); and

[0301] Its analogues, derivatives and salts;

[0302] 2) Agents that stimulate the immune system, including but not limited to:

[0303] i) Agonists / activators of tumor necrosis factor receptor superfamily member 4 (TNFRSF4, OX40 or CD134), such as OX40-targeting antibodies (e.g., MEDI-6469 and 9B12) and OX40 ligands (e.g. OX40L);

[0304] ii) Agonists / activators of TNFRSF member 5 (TNFRSF5 or CD40), such as CD40-targeting antibodies (e.g., dasizumab and CP-870, 893) and CD40 ligands (e.g., CD40L[CD154]);

[0305] iii) Agonists / activators of TNFRSF member 9 (TNFRSF9, 4-1BB or CD137), such as 4-1BB-targeting antibodies (e.g., urilumab [BMS-663513] and PF-05082566) and ligands of 4-1BB (e.g., 4-1BBL);

[0306] iv) Agonists / activators of TNFRSF member 18 (TNFRSF18, glucocorticoid-induced TNFR-associated protein [GITR] or CD357), such as GITR-targeting antibodies (e.g., DTA-1 and TRX518) and GITR ligands (e.g., GITRL).

[0307] v) Agonists / activators of Toll-like receptors (TLRs), such as ligands of TLR9 (e.g., unmethylated CpG oligodeoxynucleotides [CpG ODN], such as atomod);

[0308] vi) Cytokines and hormones that stimulate immune cells, such as interleukin-6 (IL-6) and adrenaline (e.g., stimulants of natural killer cells); and

[0309] Its analogues, derivatives, fragments, and salts;

[0310] 3) Agents that block immune checkpoints, including but not limited to:

[0311] i) Inhibitors of programmed cell death 1 (PD-1) receptor or its ligands (e.g., PD-L1 and PD-L2), such as anti-PD-1 antibodies (e.g., cimiprilmab [REGN-2810], nivolumab [BMS-936558, MDX-1106 or ONO-4538], pembrolizumab [lanolizumab or MK-3475], pildizumab [CT-011] and MEDI-0680 [AMP-514]), anti-PD-1 fusion proteins (e.g., AMP-224 [containing F... c Anti-PD-L1 antibodies (e.g., avelumumab [MSB0010718C], atezolizumab [MPDL3280A or RG7446], durvalumab [MEDI-4736], BMS-936559 [MDX-1105], LY3300054 and YW243.55.S70) and small molecule inhibitors of PD-L1 (e.g., BMS-1001 and BMS-1166);

[0312] ii) Inhibitors of the cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) receptor or its ligands, such as anti-CTLA-4 antibodies (e.g., ipilimumab and trimemumab);

[0313] iii) Inhibitors of killer cell immunoglobulin-like receptors (KIRs) or their ligands, such as anti-KIR antibodies (e.g., lirelurumab [IPH2102 or BMS-986015]);

[0314] iv) Inhibitors of the lymphocyte activation gene 3 (LAG-3) receptor or its ligands, such as anti-LAG-3 antibodies (e.g., renalalimab [BMS-986016] and GSK2831781);

[0315] v) Inhibitors of T-cell immunoglobulin and mucin domain containing 3 (TIM-3, also known as hepatitis A virus cell receptor 2 [HAVCR2]), such as anti-TIM3 antibodies (e.g., LY3321367, MBG453 and TSR-022);

[0316] vi) Inhibitors of indoleamine 2,3-dioxygenase (IDO or IDO1), such as indotimod (1-methyl-D-tryptophan or NLG-8189), navotimod (GDC-0919), α-methyl-tryptophan, β-carboline (9H-pyrido[3,4-b]indole or norhalman), icardolstat (INCB024360), BMS-986205, NLG-919, and cyclooxygenase 2 (COX-2) inhibitors (e.g., coxib [e.g., alpracoxib, celecoxib, etoricoxib, lucicoxib, parecoxib, rofecoxib, and vardicoxib], which downregulate IDO expression); and

[0317] Its analogues, derivatives, fragments, and salts;

[0318] 4) Inhibitors of angiogenesis, including but not limited to inhibitors of vascular endothelial growth factor (VEGF) (e.g., squalane and anti-VEGF antibodies such as bevacizumab) or its receptor (VEGFR) (e.g., axitinib, fruquintinib, pazopanib, regorafenib, sorafenib, and sunitinib), and inhibitors of platelet-derived growth factor (PDGF) (e.g., squalane) or its receptor (PDGFR) (e.g., axitinib, imatinib, nilotinib, pazopanib, sorafenib, and sunitinib). Inhibitors of fibroblast growth factor (FGF) (e.g., squalane) or its receptor (FGFR) (e.g., erdatinib, pazopanib, and anti-FGFR antibodies and their fragments), angiopoietin (e.g., anti-angiopoietin antibodies such as nesivamab and AMG780) or their receptors, integrin inhibitors (e.g., ALG-1001 and JSM-6427), anconitine (anecostatin acetate), angiostatin (e.g., angiostatin K1-3), α Vβ3 inhibitors (e.g., edazolidin), berberine, bleomycin, borodin, carboxytriazole, chondroitin-derived angiogenesis inhibitors (e.g., chondromodin I and troponin I), succinate, CM101, cyclopropene fatty acids (e.g., serotonin), α-difluoromethylornithine, endostatin, everolimus, fumonisin, genistein, interferon-α, interleukin-12, itraconazole, linolenic acid, matrix metalloproteinase (MMP) inhibitors (e.g., bamasstat, cimasstat, ilomasstat, mamasstat, promasstat, remmasstat, retinoid X receptor agonists (e.g., retinoids such as bexarotin), tanomasta and tetracyclines [e.g., doxycycline, ethacycline and minocycline]), 2-methoxyestradiol Pigment epithelial-derived factor (PEDF), platelet factor-4, PPAR-γ agonists (e.g., thiazolidinediones such as sitiglitazone, rosiglitazone, netoglitazone, pioglitazone, liniglitazone, rosiglitazone, and troglitazone), prolactin, sphingosine-1-phosphate inhibitors (e.g., sonectiphos), squalene, astrocytocin, angiotensin-converting steroids (e.g., tetrahydrocortisol) plus heparin, stilbene, suramin, SU5416, taquimod, tecogallan, tetrathiomolybdate, thalidomide and its derivatives (e.g., lenalidomide and pomalidomide), thiabendazole, platelet-reactive protein (e.g., platelet-reactive protein 1), TNP-470, tranilast, solanine A, and their analogues, derivatives, fragments, and salts; and

[0319] 5) Other types of antitumor / anticancer agents, including but not limited to:

[0320] i) Inhibitors of class IA phosphoinositol 3-kinase p110α (PI3K-α), such as apeliximab, bupalixib (pan-PI3K), cupanixinib (PI3K-α / δ), pitiliximab (pan-PI3K), tacelilixib, voxtalisib (pan-PI3K), GNE-477, INK-1117, PWT-33597, SF-1126 (pan-PI3K), and ZSTK-474;

[0321] ii) Drug efflux pump inhibitors, such as P-glycoprotein inhibitors (e.g., mifepristone and verapamil);

[0322] iii) Cell adhesion inhibitors, such as cimetidine;

[0323] iv) Golgi apparatus disruptors, such as Brefieldin (e.g., Brefieldin A);

[0324] v) Ionizing radiation, such as X-rays;

[0325] vi) Radiopharmaceuticals, such as I 131 -Iodide, I 131 -MIBG (meta-iodobenzylguanidine), Ra 223 -dichloride, Sm 153 -EDTMP (ethylenediaminetetramethylenephosphonic acid) and Sr 89 -chloride;

[0326] vii) Radiosensitizers for tumors / cancer cells, such as poly(ADP-ribose) polymerase (PARP) inhibitors (e.g., niraparib, olaparib, rucaparib, tapazoparib, veriparib, 4-amino-1,8-naphthodiimide, BGB290, CEP9722 and E7016), berberine and indomethacin;

[0327] viii) Agents that enhance cell survival after cytotoxic drugs or radiation therapy, such as pirfenidone-α;

[0328] ix) Vaccines, such as those that stimulate the immune system to recognize proteins produced by tumors / cancer cells and thus attack them; and

[0329] Its analogues, derivatives and salts.

[0330] In other implementations, NNMT inhibitors are used in combination with one or more anti-obesity agents and / or one or more antidiabetic drugs to treat metabolic disorders.

[0331] Anti-obesity agents include, but are not limited to:

[0332] Appetite suppressants (anorexia agonists) include amphetamine, dextroamphetamine, amphetamine, clobenzylrexate, masindole, phentermine (with or without topiramate), and lorcaserin.

[0333] Satiety promoters include ciliary neurotrophic factor (e.g., axon factor) and long-acting analogues of amylin, calcitonin, cholecystokinin (CCK), glucagon (GCG), GLP-1, gastric inhibitory peptide (GIP, also known as glucose-dependent insulinotropic peptide), leptin, gastrin (OXM), pancreatic polypeptide (PP), peptide YY (PYY), and neuropeptide Y (NPY);

[0334] Lipase inhibitors include caulerpenyne, neolistatin, erbelin A and B, lipase inhibitor, lipresstatin, orlistat, perciquinine, pancorixin AE, valyl lactone, and styracin.

[0335] Melanocortin 4 (MC4) receptor agonists, including selective MC4R agonists (e.g., AZD-2820, LY-2112688, MK-0493, PF-00446687, PG-931, PL-6983, Ro 27-3225, and THIQ [drug name]) and non-selective MC4R agonists (e.g., afanotoxin, bumenotoxin, melanotan II, mectin melanotan, and semeranovotan);

[0336] Agents that increase energy expenditure and / or fat burning include long-acting glucagon analogs, glucagon receptor agonists (e.g., NN9030), and dual GLP-1 receptor / glucagon receptor agonists (hereinafter); triiodothyronine (T3) and thyroid hormone receptor-β (THR-β) agonists (e.g., MB07344, MB07811, MGL-3196, MGL-3745, VK0214, and VK2809); fibroblast growth factor 21 (FGF21) and its analogues and derivatives (e.g., BMS-986036 [PEGylated FGF21] and BMS-986171); and mitochondrial uncoupling agents (e.g., those providing mild uncoupling, such as nitrozonide, thiazolidinediones, BAM15, 2,4-dinitrophenol [DNP] and N-(4-cyanobicyclo[2.2.2]octane-1-yl)-4-fluoro-2-[(3,3,3-trifluoropropyl)sulfonamido]benzamide [GS-3752]);

[0337] Antilipidemia agents (detailed below);

[0338] Other agents that reduce body weight and / or fat mass include GLP-1 receptor agonists (hereinafter), dual GLP-1R / GIPR agonists (hereinafter), and triple GLP-1R / GIPR / GCGR agonists (hereinafter); and

[0339] Its analogues, derivatives and salts.

[0340] Antilipidemia agents include, but are not limited to:

[0341] HMG-CoA reductase inhibitors, including statins {e.g., atorvastatin, cerivastatin, fluvastatin, mevastatin, monoclonal (e.g., monoclonal K [lovastatin]), pitavastatin, pravastatin, rosuvastatin, and simvastatin} and flavanones (e.g., naringenin);

[0342] Squalene synthase inhibitors, including lapastatin, zalagogic acid, and RPR-107393;

[0343] Fatty acid synthase inhibitors, including TVB-2640;

[0344] Acetyl-CoA carboxylase (ACC) inhibitors, including anthocyanins, amaricin, chloroacetylated biotin, cyclodimer, quizalofop-p-ethyl, firsocostat (GS-0976, NDI-010976, or ND-630), gecarbene, flupyradifon, and soraphenes (e.g., soraphen A). 1α ), 5-(tetradecoxy)-2-furanoic acid (TOFA), CP-640186, DRM-01, PF-05175157, PF-05221304, QLT-091382; 7-(4-propoxy-phenylethynyl)-3,3-dimethyl-3,4-dihydro-2H-benzo[b][1,4]dioxane-heptene; N-ethyl-N'-(3-{[4-(3,3-dimethyl-1-oxo-2 -oxa-7-azaspiro[4.5]dec-7-yl)piperidin-1-yl]-carbonyl}-1-benzothiophene-2-yl)urea; 5-(3-acetaminobut-1-ynyl)-2-(4-propoxyphenoxy)thiazole; and 1-(3-{[4-(3,3-dimethyl-1-oxo-2-oxa-7-azaspiro[4.5]dec-7-yl)piperidin-1-yl]-carbonyl}-5-(pyridin-2-yl)-2-thiophene)-3-ethylurea;

[0345] ATP citrate lyase (ACL) inhibitors, including bepedocoxel (ETC-1002), 2-furoic acid, (-)-hydroxycitric acid, BMS-303141, MEDICA-16 and SB-204990;

[0346] PPAR-α agonists include fibrates (e.g., bezafibrate, ciprofibrate, crifibrate, clofibrate, clofibrate aluminum clofibrate [clofibrate aluminum], clofibrate, etopofibrate, fenofibrate, fenofibrate, gemfibrozil, ronifibrate and propylene glycol), isoflavones (e.g., daidzein and genistein), and perfluoroalkyl acids (e.g., perfluorooctanoic acid and perfluorononanoic acid);

[0347] PPAR-δ agonists, including elafibranor (a dual PPAR-α / δ agonist), lanifibranor (a triple PPAR-α / δ / γ agonist), GFT505 (a dual PPAR-α / δ agonist), GW0742, GW501516 (a dual PPAR-β / δ agonist), sodelglitazar (GW677954), seladelpar (MBX-8025), and isoflavones (e.g., daidzein and genistein);

[0348] PPAR-γ agonists include thiazolidinediones (hereinafter), salrogleeza (a dual PPAR-α / γ agonist), IVA-337 (a tri-PPAR-α / δ / γ agonist), 4-oxo-2-thiothiazolidinones (e.g., rhodanine), berberine, magnolol, perfluorononanoic acid, cyclopentenone prostaglandins (e.g., cyclopentenone 15-deoxy-Δ-prostaglandin J2 [15d-PGJ2]), and isoflavones (e.g., daidzein and genistein).

[0349] Hepatic X receptor (LXR) agonists include endogenous ligands (such as oxosterols like 22(R)-hydroxycholesterol, 24(S)-hydroxycholesterol, 27-hydroxycholesterol, and cholesteric acid) and synthetic agonists (such as acetyl-rohanpinic acid dimer, hypocholamide, N,N-dimethyl-3β-hydroxycholesterolamide [DMHCA], BMS-852927, GW3965, and T0901317);

[0350] Retinoid X receptor (RXR) agonists, including endogenous ligands (e.g., 9-cis-retinoic acid) and synthetic agonists (e.g., bexarotin, AGN 191659, AGN 191701, AGN 192849, BMS649, LG100268, LG100754 and LGD346);

[0351] G protein-coupled bile acid receptor 1 (TGR5) agonists, including RDX-009, INT-777 and INT-767 (dual TGR5 / FXR agonists);

[0352] Triiodothyronine and thyroid hormone receptor-β agonists (above);

[0353] Hexocolokinase inhibitors (hereinafter);

[0354] Inhibitors of acyl-CoA cholesterol acyltransferase (ACAT, also known as sterol O-acyltransferase [SOAT], including ACAT1 [SOAT1] and ACAT2 [SOAT2]) include avamidib, partemidib, sennae, terpenoid C and flavanones (e.g., naringenin).

[0355] Stearoyl-CoA desaturase-1 (SCD-1, also known as stearoyl-CoA) Inhibitors of Δ9 desaturase activity or expression, including aramchol, CAY-10566, CVT-11127, SAR-224, SAR-707, XEN-103; 3-(2-hydroxyethoxy)-4-methoxy-N-[5-(3-trifluoromethylbenzyl)thiazolyl-2-yl]benzamide and 4-ethylamino-3-(2-hydroxyethoxy)-N-[5-(3-trifluoromethylbenzyl)thiazolyl-2-yl]benzamide; 1'-{6-[5-(pyridin-3-ylmethyl)-1,3,4-oxadiazol-2-yl]pyridazin-3-yl}-5-(trifluoromethyl)-3,4-dihydrospiro[chromene-2,4'-piperidine]; 5-fluoro-1'-{6-[5-(pyridin- 3-(3-ylmethyl)-1,3,4-oxadiazol-2-yl]pyridazine-3-yl}-3,4-dihydrospiro[chromene-2,4'-piperidine]; 6-[5-(cyclopropylmethyl)-4,5-dihydro-1'H,3H-spiro[1,5-benzoxazon-2,4'-piperidine]-1'-yl]-N-(2-hydroxy-2-pyridin-3-ylethyl)pyridazine-3-carboxamide; 6-[4-(2-methylbenzoyl)piperidin-1-yl]pyridazine-3-carboxylic acid (2-hydroxy-2-pyridin-3-ylethyl)amide; 4-(2-chlorophenoxy)-N-[3-(methylcarbamoyl)phenyl]piperidin-1-carboxamide; cis-9, trans-11 isomers and trans-10, cis-12 isomers of conjugated linoleic acid, WO The substituted heteroaromatic compounds, antisense polynucleotides and peptide nucleic acids (PNAs) targeting SCD-1 mRNA, and siRNAs targeting SCD-1 disclosed in 2009 / 129625 A1;

[0356] Cholesterol ester transfer protein (CETP) inhibitors, including acetrapip, dacetrapip, itettrapip, tocetrapip, and AMG 899 (TA-8995);

[0357] Inhibitors of microsomal triglyceride transfer protein (MTTP) activity or expression, including impetatape, lometatape, delotetape, miretape, CP-346086, JTT-130, SLx-4090, antisense polynucleotides and PNAs targeting MTTP mRNA, microRNAs targeting MTTP (e.g., miRNA-30c), and siRNAs targeting MTTP.

[0358] GLP-1 receptor agonists (hereinafter), glucagon receptor agonists (e.g., NN9030) and dual GLP-1 receptor / glucagon receptor agonists (hereinafter);

[0359] Inhibitors of proprotein convertase subtilisin / kexin type 9 (PCSK9) activity or expression, including berberine (which reduces PCSK9 levels), annexin A2 (which inhibits PCSK9 activity), anti-PCSK9 antibodies (e.g., aliximumab, bercocilizumab, evolocumab, LGT-209, LY3015014, and RG7652), peptides that mimic the PCSK9-binding LDL receptor epidermal growth factor-A (EGF-A) domain, PCSK9-binding adnectin (e.g., BMS-962476), antisense polynucleotides and PNAs targeting PCSK9 mRNA, and siRNAs targeting PCSK9 (e.g., incoxlane [ALN-PCS] and ALN-PCS02);

[0360] FGF21 and its analogues and derivatives (above);

[0361] Apolipoprotein mimic peptides, including apoA-I mimics (e.g., 2F, 3F, 3F-1, 3F-2, 3F-14, 4F, 4F-P-4F, 4F-IHS-4F, 4F2, 5F, 6F, 7F, 18F, 5A, 5A-C1, 5A-CH1, 5A-CH2, 5A-H1, 18A, 37pA [18A-P-18A], ELK [name], ELK-1A, ELK-1F, ELK-1K1A1E, ELK-1L1K, ELK-1W, ELK-2A, ELK-2A2K2E, ELK-2E2K, ELK-2F, etc. ELK-3E3EK, ELK-3E3K3A, ELK-3E3LK, ELK-PA, ELK-P2A, ELKA [name], ELKA-CH2, ATI-5261, CS-6253, ETC-642, FAMP [name], FREL [name] and KRES [name]) and apoE analogues (e.g., Ac-hE18A-NH2 [AEM-28], Ac-[R]hE18A-NH2, AEM-28-14, EpK, hEp, mR18L, COG-112, COG-133 and COG-1410);

[0362] Omega-3 fatty acids, including docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), alpha-linolenic acid (ALA), fish oil (which contains, for example, DHA and EPA) and their esters (e.g., glycerides and ethyl esters); and

[0363] Its analogues, derivatives and salts.

[0364] Antidiabetic drugs include, but are not limited to:

[0365] AMP-activated protein kinase (AMPK) agonists, including biguanides (e.g., bufomin, metformin and phenformin) and allosteric AMPK activators (e.g., MK-8722 and PF-793);

[0366] Peroxisome proliferator-activated receptor γ (PPAR-γ) agonists include thiazolidinediones (e.g., bagliflozin, sitiglitazone, dapagliflozin, empagliflozin, rosiglitazone, netogliflozin, pioglitazone, deuterated(R)-pioglitazone [e.g., DRX-065], liniglitazone, rosiglitazone and trogliflozin), salrogliza (a dual PPAR-α / γ agonist) and IVA-337 (a triple PPAR-α / δ / γ agonist);

[0367] Glucagon-like peptide-1 (GLP-1) receptor agonists include exenatide-4, abiglutide, dulaglutide, exenatide, liraglutide, lixilatide, smegglutide, tasglutide, tesipatide, AC-3174, CNTO736, CNTO3649, HM11260C (LAPS-Exendin), NN9926 (OG9S7GT), TT401, and ZY0G1;

[0368] Dual GLP-1 receptor (GLP-1R) / glucagon receptor (GCGR) agonists, including pemvidutide and long-acting gastric acid regulator analogs {e.g., lipid-conjugated OXM analogs (e.g., DualAG disclosed in A. Pocai et al., Diabetes, 58:2258-2266

[2009] ), PEGylated OXM analogs, cross-linked OXM analogs and R disclosed in A. Muppidi et al., ACS Chem. Biol., 11:324-328 (2016)}. The following are disclosed in Scott et al., Peptides, 104:70-77 (2018): OX-SR}, HM12525A, JNJ-54728518, LY2944876(TT-401), MEDI0382, MK-8521, MOD-6031, NN9277, SAR425899, SP-1373 and ZP2929;

[0369] Dual GLP-1R / glucose-dependent insulinotropic peptide receptor (GIPR) agonists, including Cpd86, LY3298176, NN9709 (MAR709), SAR438335, ZP-DI-70, and ZP-I-98;

[0370] Triple GLP-1R / GIPR / GCGR agonists, including HM15211 and MAR423;

[0371] Dipeptidyl peptidase-4 (DPP-4) inhibitors include alogliptin, annagliptin, dugliptin, ilagliptin, giglitazone, gogliptin, linagliptin, oxagliptin, saxagliptin, selegiline, sitagliptin, defluxigliptin, terliline, teraliline, and vidaliline;

[0372] Glucokinase activators, including pioglitazone, ARRY-403, HMS-5552, TMG-123, and TTP-399;

[0373] Alpha-glucosidase inhibitors, including acarbose, miglitol, and voglibose;

[0374] Hexokinase (KHK) inhibitors, including PF-06835919;

[0375] Sodium-glucose transporter 2 (SGLT2) inhibitors include betagliflozin, canagliflozin (which also inhibits SGLT1), dapagliflozin, empagliflozin, eleggliflozin, ioggliflozin, repaggliflozin ecaponicate, soragliflozin (which also inhibits SGLT1), and toragliflozin.

[0376] ATP-dependent K+ on pancreatic β-cells + (K ATP ) channel blockers, including meglinide (e.g. miglinide, nateglinide, and repaglinide) and sulfonylureas {including first-generation (e.g., acesulfame potassium, amiloride, chlorpropamide, glibenclamide [tolasulfonylurea], methaqualone, tolasulfonylurea, and tolbutamide) and second-generation (e.g., glibenclamide [eubrinose], glibenclamide, gliclazide, glimepiride, glipizide, gliquidone, glioxazole, and glipizide)};

[0377] Insulin and its analogues include rapid-acting insulin (e.g., insulin aspart, insulin glutares, and insulin lispro), intermediate-acting insulin (e.g., insulin NPH), and long-acting insulin (e.g., insulin degludec, insulin detemir, and insulin glargine).

[0378] Amylin and its analogues, including pramlintide; and

[0379] Its analogues, derivatives and salts. Example

[0380] The following examples are intended to illustrate this disclosure only. Other synthetic methods, assays, studies, protocols, procedures, methodologies, techniques, reagents, and conditions may be used as appropriate.

[0381] Compound Synthesis

[0382] General Synthesis Method

[0383] All air- or moisture-sensitive reactions were performed under positive nitrogen pressure using dried glassware. Anhydrous solvents such as dichloromethane (DCM), N,N-dimethylformamide (DMF), acetonitrile, methanol, and triethylamine were purchased from Sigma-Aldrich. Preparative purifications were performed on a Waters semi-preparative HPLC system. Phenomenex Luna columns were used. TM C18 column (5 μm, 30 x 75 mm), flow rate 45 mL / min. Unless otherwise specified, use Acid Standard Gradient Method Acidic Standard Gradient) (10:90 MeCN containing 0.1% trifluoroacetic acid (TFA) / deionized water containing 0.1% TFA increased to 100% deionized water containing 0.1% TFA) or Basic Standard Gradient Method Purification was performed over 8 minutes using a mixture of 10:90 MeCN containing 0.1% NH4OH / deionized water containing 0.1% NH4OH to 100% deionized water containing 0.1% NH4OH. Fraction collection was triggered by UV detection (220 nM). Analysis was performed on an Agilent LC / MS (Agilent Technologies, Santa Clara, Cal.). Purity analysis was performed using a 7-minute gradient from 4% to 100% acetonitrile (containing 0.025% TFA) in water (containing 0.05% TFA), with a run time of 8 minutes and a flow rate of 1 mL / min. Phenomenex Luna was used. TM C18 column (3 micrometers, 3 x 75 mm), temperature 50°C, using an Agilent diode array detector.

[0384] Chiral separation was performed using an Agilent 1200 series system. The column used was a Chiralpak® IG (20 µM, 5 x 50 cm), and the flow rate was 35 mL / min. The mobile phase consisted of hexane, ethanol, and diethylamine (50 / 50 / 0.05).

[0385] Mass determination was performed using an Agilent 6130 mass spectrometer with positive ion mode electrospray ionization (ESI). 1 ¹H NMR spectroscopy was recorded using a Varian 400 MHz spectrometer. Chemical shifts are reported in ppm, with a non-deuterated solvent (DMSO-d5 peak at 2.50 ppm) used as an internal standard for DMSO-d6 solution. All compounds tested in the bioassays were analyzed by LCMS with a purity greater than 95%. High-resolution mass spectrometry was recorded using an Agilent 6210 time-of-flight LC / MS system. Molecular formulas were confirmed by electrospray ionization in positive ion mode using Agilent Masshunter software (version B.02).

[0386] Example 1

[0387] The following process describes the synthesis of exemplary compounds.

[0388]

[0389] Step 1: To a solution of (S)-1-(tert-butyloxycarbonyl)piperidine-3-carboxylic acid (10 g, 43.6 mmol) in DCM (210 mL), 1-hydroxybenzotriazole (HOBt) (8.25 g, 61.1 mmol), EDC (10.03 g, 52.3 mmol), DIPEA (19.04 mL, 109 mmol), and (4-methoxyphenyl)hydrazine hydrochloride (9.14 g, 52.3 mmol) were added. The reaction mixture was stirred at room temperature for 1.5 hours. The mixture was quenched with 200 mL of water. The organic layer was dried over MgSO4 and concentrated. The crude product was purified by ISCO (30-80%, EtOAc / hexane) to give tert-butyl (S)-3-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)piperidine-1-carboxylic acid (13.5 g, 89% yield). MS m / z (M+H + 350.2.

[0390] Step 2: At 0 °C, 2,2,2-trichloroacetyl isocyanate (0.477 ml, 4.01 mmol) was added to a solution of (S)-3-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)piperidine-1-carboxylic acid tert-butyl ester (1 g, 2.86 mmol) in DCM (30 ml). The reaction mixture was stirred at 0 °C for 10 min. The solvent was evaporated. The residue was treated with MeOH (30.0 ml) and potassium carbonate (1.582 g, 11.45 mmol). The reaction mixture was stirred at room temperature for 2 h, then stirred at 65 °C for 3 days. The reaction was quenched with water and extracted with EtOAc. The organic layer was dried over MgSO4 and concentrated. The crude product was purified by ISCO (0%-50%, EtOAc / DCM) to give (S)-3-(1-(4-methoxyphenyl)-5-oxo-2,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-carboxylic acid tert-butyl ester. MS m / z (M+H + 375.1.

[0391] Step 3: Add TFA (0.226 ml, 2.94 mmol) to a solution of (S)-3-(1-(4-methoxyphenyl)-5-oxo-2,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-carboxylic acid tert-butyl ester (0.11 g, 0.294 mmol) in DCM (2 ml). Stir the mixture at room temperature for 4 hours. Evaporate the solvent. The crude product (S)-2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one was used directly for the next step without purification. MS m / z (M+H + )275.1.

[0392] Step 4: To a solution of (S)-2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (0.08 g, 0.292 mmol) in 1,2-dichloroethane (2 ml), add DIPEA (0.102 ml, 0.583 mmol), acetic acid (0.025 ml, 0.437 mmol), and 3-formylbenzamide (0.043 g, 0.292 mmol). Stir the mixture at room temperature for 30 minutes, then add sodium triacetoxyborohydride (0.124 g, 0.583 mmol). Stir the reaction mixture at room temperature for 3 hours. Quench the reaction with water and extract with EtOAc (3 times). Dry and concentrate the combined organic layers. The crude product was purified by ISCO (0-10%, MeOH / DCM) to give (S)-3-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)benzamide (compound 18).

[0393]

[0394] Synthesis of tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylate (compound 1):

[0395]

[0396] Compound 1

[0397] The title compound was prepared according to Example 1, using tert-butyl 3-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)piperidine-1-carboxylate as the starting material for step 2.

[0398]

[0399] Synthesis of 2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 2):

[0400]

[0401] Compound 2

[0402] The title compound was prepared according to Example 1, using tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylate as the starting material for step 3. MS m / z (M+H + 275.2.

[0403] Synthesis of 5-(1-benzylpiperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 3):

[0404]

[0405] Compound 3

[0406] The title compound was prepared according to Example 1, using benzaldehyde as the starting material for step 4.

[0407]

[0408] Synthesis of 2-(4-methoxyphenyl)-5-(1-(pyridin-2-ylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 6):

[0409]

[0410] Compound 6

[0411] The title compound was prepared according to Example 1, using pyridine-2-carboxaldehyde as the starting material for step 4. MS m / z (M+H) + 366.4.

[0412] Synthesis of 2-(4-methoxyphenyl)-5-(1-(pyridin-4-ylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 7):

[0413]

[0414] Compound 7

[0415] The title compound was prepared according to Example 1, using isonicoaldehyde as the starting material for step 4.

[0416]

[0417] Synthesis of 2-(4-methoxyphenyl)-5-(1-(pyridin-3-ylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 8):

[0418]

[0419] Compound 8

[0420] The title compound was prepared according to Example 1, using nicotinic aldehyde as the starting material for step 4.

[0421]

[0422] Synthesis of tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)azacyclobutane-1-carboxylate (compound 10):

[0423]

[0424] Compound 10

[0425] The title compound was prepared according to Example 1, using tert-butyl 3-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)azacyclobutane-1-carboxylate as the starting material for step 2.

[0426]

[0427] Synthesis of 5-(azacyclobutane-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 11):

[0428]

[0429] Compound 11

[0430] The title compound was prepared according to Example 1, using tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)azacyclobutane-1-carboxylate as the starting material for step 3.

[0431]

[0432] Synthesis of 2-(4-methoxyphenyl)-5-(1-(cyclohexylmethyl)azacyclobutane-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 12):

[0433]

[0434] Compound 12

[0435] The title compound was prepared from 5-(azacyclobutan-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one according to Example 1, using cyclohexaneformaldehyde as the starting material for step 4.

[0436]

[0437] Synthesis of tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)pyrrolidine-1-carboxylate (compound 13):

[0438]

[0439] Compound 13

[0440] The title compound was prepared according to Example 1, using tert-butyl 3-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)pyrrolidine-1-carboxylate as the starting material for step 2.

[0441]

[0442] Synthesis of 2-(4-methoxyphenyl)-5-(pyrrolidone-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 14):

[0443]

[0444] Compound 14

[0445] The title compound was prepared according to Example 1, using tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-2,5-dihydro-1H-1,2,4-triazol-3-yl)pyrrolidine-1-carboxylate as the starting material for step 3.

[0446]

[0447] Synthesis of 5-(1-(cyclohexylmethyl)pyrrolidone-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 15):

[0448]

[0449] Compound 15

[0450] The title compound was prepared from tert-butyl 3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)pyrrolidine-1-carboxylate according to Example 1, using cyclohexaneformaldehyde as the starting material for step 4.

[0451]

[0452] Synthesis of 2-(4-methoxyphenyl)-5-((3S)-1-((tetrahydro-2H-pyran-3-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 17):

[0453]

[0454] Compound 17

[0455] The title compound was prepared from (S)-2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one according to Example 1, using tetrahydro-2H-pyran-3-carboxaldehyde as the starting material for step 4.

[0456]

[0457] Synthesis of 2-(4-methoxyphenyl)-5-((3S)-1-((tetrahydro-2H-pyran-2-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 23):

[0458]

[0459] Compound 23

[0460] The title compound was prepared from (S)-2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one according to Example 1, using tetrahydro-2H-pyran-2-carboxaldehyde as the starting material for step 4. MS m / z (M+H + 373.1.

[0461] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(pyridin-2-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 24):

[0462]

[0463] Compound 24

[0464] The title compound was prepared according to Example 1, using 2-hydrazinopyridine as the starting material in step 1 and cyclohexaneformaldehyde as the starting material in step 4.

[0465]

[0466] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(pyridin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 25):

[0467]

[0468] Compound 25

[0469] The title compound was prepared according to Example 1, using 3-hydrazinopyridine as the starting material in step 1 and cyclohexanecarboxaldehyde as the starting material in step 4. MS m / z (M+H) + 342.2.

[0470] Synthesis of 5-(piperidin-3-yl)-2-(pyridin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 26):

[0471]

[0472] Compound 26

[0473] The title compound was prepared according to Example 1, using tert-butyl 3-(5-oxo-1-(pyridin-3-yl)-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylate as the starting material for step 3.

[0474]

[0475] Synthesis of (S)-5-(1-(3-cyclohexylpropyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 92):

[0476]

[0477] Compound 92

[0478] The title compound was prepared according to Example 1, using 3-cyclohexylpropionaldehyde (0.051 g, 0.365 mmol) as the starting material for step 4.

[0479]

[0480] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((5-methylpyrimidin-2-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 100):

[0481]

[0482] Compound 100

[0483] The title compound was prepared according to Example 1, using 5-methylpyrimidine-2-carboxaldehyde as the starting material for step 4.

[0484]

[0485] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(pyrimidin-2-ylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 101):

[0486]

[0487] Compound 101

[0488] The title compound was prepared according to Example 1, using pyrimidine-2-carboxaldehyde as the starting material for step 4.

[0489]

[0490] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((4-methylpyrimidin-2-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 102):

[0491]

[0492] Compound 102

[0493] The title compound was prepared according to Example 1, using 4-methylpyrimidine-2-carboxaldehyde as the starting material for step 4.

[0494]

[0495] Synthesis of (S)-2-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)isonicotinic acid methyl ester, TFA (compound 104):

[0496]

[0497] Compound 104

[0498] The title compound was prepared according to Example 1, using methyl 2-(bromomethyl)isonicotinic acid as the starting material for step 4.

[0499]

[0500] Synthesis of (S)-3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylic acid tert-butyl ester (compound 128):

[0501]

[0502] Compound 128

[0503] The title compound was prepared according to Example 1, using 5-hydrazinoisoquinoline as the starting material for step 1.

[0504]

[0505] Synthesis of (S)-2-(isoquinolin-5-yl)-5-(1-(2-(naphthyl-2-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 134):

[0506]

[0507] Compound 134

[0508] The title compound was prepared according to Example 1, using 5-hydrazinoisoquinoline in step 1 and 2-(naphth-2-yl)acetaldehyde as the starting material in step 4.

[0509]

[0510] Synthesis of (S)-2-(isoquinolin-5-yl)-5-(1-(2-(naphthyl-1-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 136):

[0511]

[0512] Compound 136

[0513] The title compound was prepared according to Example 1, using 5-hydrazinoisoquinoline in step 1 and 2-(naphth-1-yl)acetaldehyde as the starting material in step 4.

[0514]

[0515] Example 1a

[0516] The following process describes the synthesis of exemplary compounds.

[0517]

[0518] Step 1: Tert-butyl carbamate ((1R,4S)-4-(2-((S)-3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)ethyl)cyclohexyl)carbamate was prepared according to Example 1, using tert-butyl carbamate ((1S,4S)-4-(2-oxoethyl)cyclohexyl)carbamate as the starting material for Step 4. MS m / z (M+H + 500.3.

[0519] Step 2: Add TFA (0.2 ml, 2.60 mmol) to a solution of ((1R,4S)-4-(2-(((S)-3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)ethyl)cyclohexyl)carbamate tert-butyl ester (0.18 g, 0.360 mmol) in DCM (1 ml). Stir the mixture at room temperature for a period of time. Remove the solvent. Submit the crude product for purification to give 5-((S)-1-(2-(((1S,4R)-4-aminocyclohexyl)ethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, 2 TFA (compound 94).

[0520]

[0521] Synthesis of tert-butyl piperidine-1-carboxylate (compound 88): 3-(((S)-3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)piperidin-1-carboxylate

[0522]

[0523] Compound 88

[0524] The title compound was prepared according to Example 1a, using tert-butyl 3-formylpiperidine-1-carboxylate as the starting material for step 1.

[0525]

[0526] Synthesis of 3-(((S)-3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)piperidine (compound 90):

[0527]

[0528] Compound 90

[0529] The title compound was prepared according to Example 1a, using tert-butyl 3-(((S)-3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)piperidin-1-carboxylic acid as the starting material for step 2.

[0530]

[0531] Synthesis of 5-((S)-1-(2-((1R,4S)-4-aminocyclohexyl)ethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, 2 TFA (compound 93):

[0532]

[0533] Compound 93

[0534] The title compound was prepared according to Example 1a, using ((1R,4R)-4-(2-oxoethyl)cyclohexyl)carbamate tert-butyl ester as the starting material for step 1.

[0535]

[0536] Example 1b

[0537] The following process describes the synthesis of exemplary compounds.

[0538]

[0539] Trifluoroacetic acid (267 µl, 3.47 mmol) was added to a solution of (S)-3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylic acid tert-butyl ester (130 mg, 0.347 mmol) in CH2Cl2 (1 mL). The reaction was stirred at room temperature for 1.5 h. The solvent was removed, and the residue was dried under high vacuum for 2 h. The residue was dissolved in DMF (1 mL) and treated with a suspension of potassium carbonate (240 mg, 1.736 mmol) and tetrahydro-2H-pyran-4-yl)methanesulfonate (81 mg, 0.417 mmol) in DMF (0.5 mL). The reaction mixture was stirred at room temperature for 3 h, and then heated at 50 °C for 3 days. The reaction was quenched with water and extracted with 10% iPrOH / CH2Cl2. The combined organic layers were washed with water and brine and concentrated. The residue was purified by rapid chromatography to give (S)-2-(4-methoxyphenyl)-5-(1-((tetrahydro-2H-pyran-4-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 16).

[0540]

[0541] Example 1c

[0542] The following process describes the synthesis of exemplary compounds.

[0543]

[0544] Step 1: To a mixture of 4-(tert-butoxycarbonyl)morpholine-2-carboxylic acid (5.0 g, 21.62 mmol) and 1-hydroxybenzotriazole hydrate (HOBt, 4.09 g, 30.3 mmol) in DMF (15 mL), EDC HCl (4.97 g, 25.9 mmol), triethylamine (7.53 mL, 54.1 mmol), and (4-methoxyphenyl)hydrazine hydrochloride (4.53 g, 25.9 mmol) were added. The reaction mixture was stirred at room temperature for 24 hours. The reaction mixture was filtered through a porous glass funnel. The filtrate was diluted with water (150 mL) and extracted with EtOAc (3 × 200 mL). The combined organic layers were washed with water and brine and concentrated. The residue was purified by rapid chromatography. The fractions eluted with 80% EtOAc / hexane were combined and concentrated to give tert-butyl 2-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)morpholine-4-carboxylate (4.988 g, 14.19 mmol, yield 65.6%), which was a white powder.

[0545] Step 2: At 0 °C, 2,2,2-trichloroacetyl isocyanate (0.779 mL, 6.53 mmol) was added to a solution of 2-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)morpholine-4-carboxylate (1.64 g, 4.67 mmol) in CH2Cl2 (30 mL). The reaction was stirred at 0 °C for 30 min. The solvent was evaporated. The residue was treated with MeOH (20 mL) and potassium carbonate (2.58 g, 18.67 mmol). The reaction was stirred at room temperature for 3.5 h, then at 65 °C for 3 days. The solvent was removed, and the residue was purified by rapid chromatography. The fractions eluted with EtOAc were combined and concentrated to give 2-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)morpholine-4-carboxylate (compound 131).

[0546]

[0547] Step 3: Trifluoroacetic acid (614 µl, 7.97 mmol) was added to a solution of 2-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)morpholine-4-carboxylic acid tert-butyl ester (250 mg, 0.664 mmol) in CH2Cl2 (1 mL). The reaction mixture was stirred for 2.5 days. The solvent was removed, and the residue was dried overnight under high vacuum. The residue was redissolved in 1,2-dichloroethane / THF (2 mL: 1 mL). Hunig base (347 µl, 1.992 mmol) was added, followed by 2-cyclohexylacetaldehyde (168 µl, 1.328 mmol). Sodium triacetoxyborohydride (282 mg, 1.328 mmol) was then added. The reaction mixture was stirred for 1 hour. The reaction was quenched with 1 N NaOH solution (0.5 mL). After removing the solvent, the residue was purified by rapid chromatography to give 5-(4-(2-cyclohexylethyl)morpholin-2-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 137).

[0548]

[0549] Example 1d

[0550] The following process describes the synthesis of exemplary compounds.

[0551]

[0552] Step 1: To a mixture of 1,4-bis(tert-butyloxycarbonyl)piperazine-2-carboxylic acid (1.0 g, 3.03 mmol) and 1-hydroxybenzotriazole hydrate (0.573 g, 4.24 mmol) in DMF (20 mL), EDC HCl (0.696 g, 3.63 mmol), Hunig base (1.322 mL, 7.57 mmol), and (4-methoxyphenyl)hydrazine hydrochloride (0.634 g, 3.63 mmol) were added. The reaction mixture was stirred at room temperature for 20 hours. The reaction mixture was quenched with water. The mixture was filtered through a funnel. The filtrate was collected and purified by rapid chromatography. The fractions eluted with 60% EtOAc / hexane were combined and concentrated to give di-tert-butyl 2-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)piperazine-1,4-dicarboxylate (540 mg, 1.199 mmol, yield 39.6%), which was a light yellow powder.

[0553] Step 2: At 0 °C, 2,2,2-trichloroacetyl isocyanate (200 µl, 1.678 mmol) was added to a solution of di-tert-butyl 2-(2-(4-methoxyphenyl)hydrazine-1-carbonyl)piperazine-1,4-dicarboxylate (540 mg, 1.199 mmol) in CH2Cl2 (20 mL). The reaction was stirred at 0 °C for 1 hour. The solvent was evaporated. The remaining crude product was dissolved in MeOH (15 mL) and treated with potassium carbonate (663 mg, 4.79 mmol). The reaction was stirred at room temperature for 20 hours, and then the reaction mixture was heated at 60 °C for 6 days. The reaction mixture was concentrated and purified by rapid chromatography. The fractions eluted near 80% EtOAc / hexane were collected and combined to give di-tert-butyl 2-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperazine-1,4-dicarboxylate (40 mg, 0.084 mmol, yield 7.02%), as a pale yellow oil.

[0554] Step 3: Add TFA (405 µl, 5.26 mmol) to a solution of 2-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperazine-1,4-dicarboxylic acid di-tert-butyl ester (25 mg, 0.053 mmol) in CH2Cl2 (1 mL). Stir the reaction mixture at room temperature for 30 minutes. Evaporate the solvent and dissolve the residue in THF (1 mL). Add Hunig base (23.42 µl, 0.134 mmol), 2-cyclohexylacetaldehyde (8.85 µl, 0.064 mmol), and sodium triacetoxyborohydride (17.05 mg, 0.080 mmol). Stir the reaction mixture as a slurry for 40 minutes. Quench the reaction with a few drops of 1N NaOH solution. After removing the solvent, the crude product was purified by rapid chromatography to give two products: 5-(4-(2-cyclohexylethyl)piperazin-2-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 292), MS m / z (M+H + ) 386.3, and 5-(1-(2-cyclohexylethyl)piperazin-2-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 293), MS m / z (M+H + 386.3.

[0555]

[0556] Compound 292 Compound 293

[0557] Example 2

[0558] The following process describes the synthesis of exemplary compounds.

[0559]

[0560] Potassium carbonate (126 mg, 0.911 mmol) and 4-(bromomethyl)benzamide (46.8 mg, 0.219 mmol) were added to a solution of (S)-2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)benzamide (compound 19) in DMF (2 ml) to which a solution of (S)-2-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)benzamide was added. The reaction mixture was stirred at room temperature for 30 min. The crude product was purified by ISCO (0-20%, MeOH / DCM) to give (S)-4-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)benzamide.

[0561]

[0562] Synthesis of 5-(1-(cyclopropylmethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 9):

[0563]

[0564] Compound 9

[0565] The title compound was prepared according to Example 2, using (bromomethyl)cyclopropane as the starting material. MS m / z (M+H) + 329.4.

[0566] Synthesis of (S)-N-(3-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)phenyl)acetamide (compound 20):

[0567]

[0568] Compound 20

[0569] The title compound was prepared according to Example 2, using N-(3-(bromomethyl)phenyl)acetamide as the starting material.

[0570]

[0571] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-phenylethylpiperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 21):

[0572]

[0573] Compound 21

[0574] The title compound was prepared according to Example 2, using (2-bromoethyl)benzene as the starting material.

[0575]

[0576] Synthesis of (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 22):

[0577]

[0578] Compound 22

[0579] The title compound was prepared according to Example 2, using (2-bromoethyl)cyclohexane as the starting material.

[0580]

[0581] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((6-methylpyridin-2-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 30):

[0582]

[0583] Compound 30

[0584] The title compound was prepared according to Example 2, using 2-(bromomethyl)-6-methylpyridine as the starting material.

[0585]

[0586] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(4-(trifluoromethoxy)benzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 31):

[0587]

[0588] Compound 31

[0589] The title compound was prepared according to Example 2, using 1-(bromomethyl)-4-(trifluoromethoxy)benzene as the starting material.

[0590]

[0591] Synthesis of (S)-5-(1-(3-methoxybenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 32):

[0592]

[0593] Compound 32

[0594] The title compound was prepared according to Example 2, using 1-(bromomethyl)-3-methoxybenzene as the starting material.

[0595]

[0596] Synthesis of (S)-5-(1-(3-fluorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 33):

[0597]

[0598] Compound 33

[0599] The title compound was prepared according to Example 2, using 1-(bromomethyl)-3-fluorobenzene as the starting material.

[0600]

[0601] Synthesis of (S)-5-(1-(3-chlorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 34):

[0602]

[0603] Compound 34

[0604] The title compound was prepared according to Example 2, using 1-(bromomethyl)-3-chlorobenzene as the starting material.

[0605]

[0606] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(4-(trifluoromethyl)benzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 35):

[0607]

[0608] Compound 35

[0609] The title compound was prepared according to Example 2, using 1-(bromomethyl)-4-(trifluoromethyl)benzene as the starting material.

[0610]

[0611] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(3-methylbenzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 36):

[0612]

[0613] Compound 36

[0614] The title compound was prepared according to Example 2, using 1-(bromomethyl)-3-methylbenzene as the starting material.

[0615]

[0616] Synthesis of (S)-5-(1-(4-isopropylbenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 37):

[0617]

[0618] Compound 37

[0619] The title compound was prepared according to Example 2, using 1-(bromomethyl)-4-isopropylbenzene as the starting material.

[0620]

[0621] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(4-((trifluoromethyl)thio)benzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 38):

[0622]

[0623] Compound 38

[0624] The title compound was prepared according to Example 2, using (4-(bromomethyl)phenyl)(trifluoromethyl) sulfide as the starting material.

[0625]

[0626] Synthesis of (S)-5-(1-(4-bromobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 39):

[0627]

[0628] Compound 39

[0629] The title compound was prepared according to Example 2, using 4-bromo-1-(bromomethyl)benzene as the starting material.

[0630]

[0631] Synthesis of (S)-N-(4-(2-(3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)ethyl)phenyl)acetamide, TFA (compound 40):

[0632]

[0633] Compound 40

[0634] The title compound was prepared according to Example 2, using N-(4-(2-bromoethyl)phenyl)acetamide as the starting material.

[0635]

[0636] Synthesis of (S)-5-(1-([1,1'-biphenyl]-4-ylmethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 41):

[0637]

[0638] Compound 41

[0639] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1,1'-biphenyl as the starting material.

[0640]

[0641] Synthesis of (S)-5-(1-(2,6-dichlorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 42):

[0642]

[0643] Compound 42

[0644] The title compound was prepared according to Example 2, using 2-(bromomethyl)-1,3-dichlorobenzene as the starting material.

[0645]

[0646] Synthesis of (S)-5-(1-(3,4-difluorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 43):

[0647]

[0648] Compound 43

[0649] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1,2-difluorobenzene as the starting material.

[0650]

[0651] Synthesis of (S)-N-(3-(2-(3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)ethyl)phenyl)acetamide, TFA (compound 44):

[0652]

[0653] Compound 44

[0654] The title compound was prepared according to Example 2, using N-(3-(2-bromoethyl)phenyl)acetamide as the starting material.

[0655]

[0656] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(3-(trifluoromethyl)benzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 45):

[0657]

[0658] Compound 45

[0659] The title compound was prepared according to Example 2, using 1-(bromomethyl)-3-(trifluoromethyl)benzene as the starting material.

[0660]

[0661] Synthesis of (S)-5-(1-(3,4-dichlorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 46):

[0662]

[0663] Compound 46

[0664] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1,2-dichlorobenzene as the starting material.

[0665]

[0666] Synthesis of (S)-5-(1-(3-(2H-tetrazol-5-yl)benzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 47):

[0667]

[0668] Compound 47

[0669] The title compound was prepared according to Example 2, using 5-(3-(bromomethyl)phenyl)-2H-tetrazole as the starting material.

[0670]

[0671] Synthesis of (S)-5-(1-(3-chloro-4-(trifluoromethyl)benzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 48):

[0672]

[0673] Compound 48

[0674] The title compound was prepared according to Example 2, using 4-(bromomethyl)-2-chloro-1-(trifluoromethyl)benzene as the starting material.

[0675]

[0676] Synthesis of (S)-5-(1-(2-fluorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 49):

[0677]

[0678] Compound 49

[0679] The title compound was prepared according to Example 2, using 1-(bromomethyl)-2-fluorobenzene as the starting material.

[0680]

[0681] Synthesis of (S)-5-(1-(4-chloro-3-fluorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 50):

[0682]

[0683] Compound 50

[0684] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1-chloro-2-fluorobenzene as the starting material.

[0685]

[0686] Synthesis of (S)-5-(1-(4-chlorobenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 51):

[0687]

[0688] Compound 51

[0689] The title compound was prepared according to Example 2, using 1-(bromomethyl)-4-chlorobenzene as the starting material.

[0690]

[0691] Synthesis of (S)-5-(1-(benzo[d][1,3]dioxacyclopenten-5-ylmethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 52):

[0692]

[0693] Compound 52

[0694] The title compound was prepared according to Example 2, using 5-(bromomethyl)benzo[d][1,3]dioxane as the starting material.

[0695]

[0696] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(4-(methanesulfonyl)benzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 53):

[0697]

[0698] Compound 53

[0699] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1-(methylsulfonyl)benzene as the starting material.

[0700]

[0701] Synthesis of (S)-5-(1-(4-fluoro-3-methylbenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 54):

[0702]

[0703] Compound 54

[0704] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1-fluoro-2-methylbenzene as the starting material.

[0705]

[0706] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(4-methyl-3-(trifluoromethyl)benzyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 55):

[0707]

[0708] Compound 55

[0709] The title compound was prepared according to Example 2, using 4-(bromomethyl)-1-methyl-2-(trifluoromethyl)benzene as the starting material.

[0710]

[0711] Synthesis of (S)-3-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)benzoic acid, TFA (compound 56):

[0712]

[0713] Compound 56

[0714] The title compound was prepared according to Example 2, using 3-(bromomethyl)benzoic acid as the starting material.

[0715]

[0716] Synthesis of (S)-5-(1-(4-(2H-tetrazol-5-yl)benzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 57):

[0717]

[0718] Compound 57

[0719] The title compound was prepared according to Example 2, using 5-(4-(bromomethyl)phenyl)-2H-tetrazole as the starting material.

[0720]

[0721] Synthesis of (S)-5-(1-(3,5-dimethylbenzyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 58):

[0722]

[0723] Compound 58

[0724] The title compound was prepared according to Example 2, using 1-(bromomethyl)-3,5-dimethylbenzene as the starting material.

[0725]

[0726] Synthesis of (S)-4-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)-N,N-dimethylbenzenesulfonamide, TFA (compound 59):

[0727]

[0728] Compound 59

[0729] The title compound was prepared according to Example 2, using 4-(bromomethyl)-N,N-dimethylbenzenesulfonamide as the starting material.

[0730]

[0731] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(2-(tetrahydro-2H-pyran-4-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 60):

[0732]

[0733] Compound 60

[0734] The title compound was prepared according to Example 2, using 4-(2-bromoethyl)tetrahydro-2H-pyran as the starting material.

[0735]

[0736] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(2-morpholinoethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 61):

[0737]

[0738] Compound 61

[0739] The title compound was prepared according to Example 2, using 4-(2-bromoethyl)morpholine and HCl as starting materials.

[0740]

[0741] Synthesis of (S)-N-(4-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)phenyl)acetamide, TFA (compound 63):

[0742]

[0743] Compound 63

[0744] The title compound was prepared according to Example 2, using N-(4-(bromomethyl)phenyl)acetamide as the starting material.

[0745]

[0746] Synthesis of (S)-5-(1-((5-fluoropyridin-3-yl)methyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 64):

[0747]

[0748] Compound 64

[0749] The title compound was prepared according to Example 2, using 3-(bromomethyl)-5-fluoropyridine as the starting material.

[0750]

[0751] Synthesis of (S)-4-(2-(3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)ethyl)benzoate, TFA (compound 65):

[0752]

[0753] Compound 65

[0754] The title compound was prepared according to Example 2, using methyl 4-(2-bromoethyl)benzoate as the starting material.

[0755]

[0756] Synthesis of (S)-5-(1-(4-methoxyphenylethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 66):

[0757]

[0758] Compound 66

[0759] The title compound was prepared according to Example 2, using 1-(2-bromoethyl)-4-methoxybenzene as the starting material.

[0760]

[0761] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((5-methylpyridin-3-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 67):

[0762]

[0763] Compound 67

[0764] The title compound was prepared according to Example 2, using 3-(bromomethyl)-5-methylpyridine as the starting material.

[0765]

[0766] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(4-(trifluoromethyl)phenethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 68):

[0767]

[0768] Compound 68

[0769] The title compound was prepared according to Example 2, using 1-(2-bromoethyl)-4-(trifluoromethyl)benzene as the starting material.

[0770]

[0771] Synthesis of (S)-5-(1-(2,4-dichlorophenylethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 69):

[0772]

[0773] Compound 69

[0774] The title compound was prepared according to Example 2, using 1-(2-bromoethyl)-2,4-dichlorobenzene as the starting material.

[0775]

[0776] Synthesis of (S)-2-(4-bromophenyl)-5-(1-(cyclohexylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 70):

[0777]

[0778] Compound 70

[0779] The title compound was prepared according to Example 1 using (4-bromophenyl)hydrazine hydrochloride as the starting material for step 1, and according to Example 2 using (bromomethyl)cyclohexane as the starting material.

[0780]

[0781] Synthesis of (S)-2-(4-bromophenyl)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 71):

[0782]

[0783] Compound 71

[0784] The title compound was prepared according to Example 1 using (4-bromophenyl)hydrazine hydrochloride as the starting material for step 1, and according to Example 2 using (2-bromoethyl)cyclohexane as the starting material.

[0785]

[0786] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(2-(pyridin-2-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 77):

[0787]

[0788] Compound 77

[0789] The title compound was prepared according to Example 2, using 2-(2-bromoethyl)pyridine as the starting material.

[0790]

[0791] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(2-(pyridin-3-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 78):

[0792]

[0793] Compound 78

[0794] The title compound was prepared according to Example 2, using 3-(2-bromoethyl)pyridine as the starting material.

[0795]

[0796] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(2-(pyridin-4-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 79):

[0797]

[0798] Compound 79

[0799] The title compound was prepared according to Example 2, using 4-(2-bromoethyl)pyridine as the starting material.

[0800]

[0801] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((5-methoxypyridin-3-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 81):

[0802]

[0803] Compound 81

[0804] The title compound was prepared according to Example 2, using 3-(bromomethyl)-5-methoxypyridine as the starting material.

[0805]

[0806] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((4-methoxypyridin-2-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 82):

[0807]

[0808] Compound 82

[0809] The title compound was prepared according to Example 2, using 2-(bromomethyl)-4-methoxypyridine as the starting material.

[0810]

[0811] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(3-phenylpropyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 83):

[0812]

[0813] Compound 83

[0814] The title compound was prepared according to Example 2, using (3-bromopropyl)benzene as the starting material.

[0815]

[0816] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-(2-(pyrimidin-5-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 95):

[0817]

[0818] Compound 95

[0819] The title compound was prepared according to Example 2, using 5-(2-bromoethyl)pyrimidine as the starting material.

[0820]

[0821] Synthesis of (S)-2-(4-methoxyphenyl)-5-(1-((4-methylpyridin-2-yl)methyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 96):

[0822]

[0823] Compound 96

[0824] The title compound was prepared according to Example 2, using 2-(bromomethyl)-4-methylpyridine as the starting material.

[0825]

[0826] Synthesis of (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 97):

[0827]

[0828] Compound 97

[0829] The title compound was prepared according to Example 1 using 5-hydrazinoisoquinoline as the starting material for step 1, and according to Example 2 using (2-bromoethyl)cyclohexane as the starting material.

[0830]

[0831] Synthesis of (S)-2-(isoquinolin-5-yl)-5-(1-(2-(pyridin-2-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 133):

[0832]

[0833] Compound 133

[0834] The title compound was prepared according to Example 1 using 5-hydrazinoisoquinoline as the starting material for step 1, and according to Example 2 using 2-(2-bromoethyl)pyridine as the starting material.

[0835]

[0836] Synthesis of (S)-5-(1-(2-(4,6-dichloropyridin-3-yl)ethyl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 135):

[0837]

[0838] Compound 135

[0839] The title compound was prepared according to Example 1 using 5-hydrazinoisoquinoline as the starting material for step 1, and according to Example 2 using 5-(2-bromoethyl)-2,4-dichloropyridine as the starting material.

[0840]

[0841] Synthesis of (S)-5-(1-(2-(4-chloropyridin-2-yl)ethyl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 141):

[0842]

[0843] Compound 141

[0844] The title compound was prepared according to Example 1 using 5-hydrazinoisoquinoline as the starting material for step 1, and according to Example 2 using 2-(2-bromoethyl)-4-chloropyridine as the starting material. MS m / z (M+H + 435.2.

[0845] Synthesis of 5-(1-(2-cyclohexylethyl)-5,5-difluoropiperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 147):

[0846]

[0847] Compound 147

[0848] The title compound was prepared according to Example 1 using 1-(tert-butoxycarbonyl)-5,5-difluoropiperidine-3-carboxylic acid and 5-hydrazinoisoquinoline as starting materials for step 1, and according to Example 2 using (2-bromoethyl)cyclohexane as starting material.

[0849]

[0850] Example 2a

[0851] The following process describes the synthesis of exemplary compounds.

[0852]

[0853] LiOH (0.5 M, 331 µl, 0.165 mmol) was added to a solution of (S)-2-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)isonicotinic acid (35 mg, 0.083 mmol, prepared according to Example 2 using 2-(bromomethyl)isonicotinic acid as a starting material) in THF (0.8 mL). The mixture was stirred at room temperature for 4 hours. The solvent was evaporated under vacuum. The crude product was submitted for purification to give (S)-2-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)isonicotinic acid, TFA (compound 105).

[0854]

[0855] Example 2b

[0856] The following process describes the synthesis of exemplary compounds.

[0857]

[0858] Methyl isonicotinamide (S)-2-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)isonicotinamide (140 mg, 0.331 mmol) was dissolved in 7N ammonia-methanol solution (3 mL, 21.00 mmol). The solution was stirred overnight at 60 °C in a sealed vial. After the reaction was complete, the solvent was removed. The crude product was purified by ISCO (0-20%, MeOH / DCM) to give (S)-2-((3-(1-(4-methoxyphenyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)methyl)isonicotinamide (compound 121).

[0859]

[0860] Example 3

[0861] The following process describes the synthesis of exemplary compounds.

[0862]

[0863] To a solution of (S)-2-(4-methoxyphenyl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (50 mg, 0.182 mmol) in DCM (2 mL), DIPEA (0.127 mL, 0.729 mmol) and cyclohexanesulfonyl chloride (0.031 mL, 0.219 mmol) were added. The mixture was stirred at room temperature for 3 hours. The reaction was quenched with water and extracted with EtOAc (3 times). The combined organic layers were dried and concentrated. The crude product was purified by ISCO (0-80%, EtOAc / hexane) to give (S)-5-(1-(cyclohexylsulfonyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 27).

[0864]

[0865] Example 4

[0866] The following process describes the synthesis of exemplary compounds.

[0867]

[0868] A mixture of (S)-2-(4-bromophenyl)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (50 mg, 0.115 mmol, prepared according to Example 1 using (4-bromophenyl)hydrazine•HCl as the starting material for step 1, and according to Example 2 using (2-bromoethyl)cyclohexane as the starting material) and copper iodide (I) (21.97 mg, 0.115 mmol) in 2-(pyrrolidine-1-yl)ethanol (1 ml, 8.50 mmol) was stirred at 160 °C for 2 days. EtOAc was added to the mixture, and the mixture was washed with water. The organic layer was dried over MgSO4 and concentrated. The crude product was purified by ISCO (0-20%, MeOH / DCM) to give (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(4-(2-(pyrrolidin-1-yl)ethoxy)phenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 75).

[0869]

[0870] Synthesis of (S)-5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(4-(2-(pyrrolidin-1-yl)ethoxy)phenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 76):

[0871]

[0872] Compound 76

[0873] The title compound was prepared according to Example 1 using (4-bromophenyl)hydrazine•HCl as the starting material for step 1, according to Example 2 using (bromomethyl)cyclohexane, and according to Example 4.

[0874]

[0875] Example 5

[0876] The following process describes the synthesis of exemplary compounds.

[0877]

[0878] Step 1: Potassium carbonate (0.338 g, 2.443 mmol) and (2-bromoethyl)cyclohexane (0.084 ml, 0.537 mmol) were added to a solution of 5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one hydrochloride (0.1 g, 0.489 mmol) in DMF (5 ml). The reaction mixture was stirred at room temperature for 30 minutes. Water was added to the mixture, and the mixture was extracted with EtOAc. The organic layer was washed with NaHCO3 (3 times), dried over MgSO4, and concentrated. The crude product 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one was used directly for the next step without further purification. MS m / z (M+H + )279.1.

[0879] Step 2: Under N2, 1-iodo-3-methoxybenzene (42.8 µl, 0.359 mmol) was added to a suspension of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (50 mg, 0.180 mmol), copper iodide (I) (6.84 mg, 0.036 mmol), and potassium carbonate (124 mg, 0.898 mmol) in DMF (2 mL), followed by (1R,2R)-N,N'-dimethyl-cyclohexane-1,2-diamine (11.33 µl, 0.072 mmol). The reaction mixture was stirred overnight at 100 °C. The reaction mixture was cooled to room temperature, quenched with water, and extracted with EtOAc (3 times). The combined organic layers were washed with saturated NaHCO3, dried over MgSO4, and concentrated. The crude product was purified to give 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(3-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 85).

[0880]

[0881] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 86):

[0882]

[0883] Compound 86

[0884] The title compound was prepared according to Example 5, using 5-iodoisoquinoline as the starting material for step 2.

[0885]

[0886] Synthesis of (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 97):

[0887]

[0888] Compound 97

[0889] The title compound was prepared according to Example 5, using 5-iodoisoquinoline as the starting material in step 2, followed by chiral separation.

[0890]

[0891] Synthesis of (R)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(isoquinolin-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (98):

[0892]

[0893] Compound 98

[0894] The title compound was prepared according to Example 5, using 5-iodoisoquinoline as the starting material in step 2, followed by chiral separation.

[0895]

[0896] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(naphth-1-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 106):

[0897]

[0898] Compound 106

[0899] The title compound was prepared according to Example 5, using 1-iodonaphthalene as the starting material for step 2. MS m / z (M+H) + )405.2.

[0900] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-phenyl-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 107):

[0901]

[0902] Compound 107

[0903] The title compound was prepared according to Example 5, using iodobenzene as the starting material for step 2.

[0904]

[0905] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2,3-dihydrobenzo[b][1,4]dioxane-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 108):

[0906]

[0907] Compound 108

[0908] The title compound was prepared according to Example 5, using 6-iodo-2,3-dihydrobenzo[b][1,4]dioxane as the starting material for step 2.

[0909]

[0910] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-4-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 110):

[0911]

[0912] Compound 110

[0913] The title compound was prepared according to Example 5, using 5-(piperidin-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one hydrochloride in step 1 and 5-iodoisoquinoline as the starting material in step 2.

[0914]

[0915] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-4-yl)-2-(2,3-dihydrobenzo[b][1,4]dioxane-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 114):

[0916]

[0917] Compound 114

[0918] The title compound was prepared according to Example 5, using the hydrochloride of 5-(piperidin-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one in step 1 and 6-iodo-2,3-dihydrobenzo[b][1,4]dioxane as the starting material in step 2.

[0919]

[0920] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(8-methoxyisoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 115):

[0921]

[0922] Compound 115

[0923] The title compound was prepared according to Example 5, using 5-bromo-8-methoxyisoquinoline as the starting material for step 2.

[0924]

[0925] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(isoquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 116):

[0926]

[0927] Compound 116

[0928] The title compound was prepared according to Example 5, using 8-iodoisoquinoline as the starting material for step 2.

[0929]

[0930] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2-methoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 117):

[0931]

[0932] Compound 117

[0933] The title compound was prepared according to Example 5, using 8-bromo-2-methoxyquinoline as the starting material for step 2.

[0934]

[0935] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-4-yl)-2-(8-methoxyisoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 118):

[0936]

[0937] Compound 118

[0938] The title compound was prepared according to Example 5, using 5-(piperidin-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one hydrochloride in step 1 and 5-bromo-8-methoxyisoquinoline as the starting material in step 2.

[0939]

[0940] Synthesis of 2-(1-aminoisoquinoline-5-yl)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 120):

[0941]

[0942] Compound 120

[0943] The title compound was prepared according to Example 5, using 5-bromoisoquinoline-1-amine as the starting material for step 2. MSm / z (M+H + 421.3.

[0944] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(6-methylisoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 122):

[0945]

[0946] Compound 122

[0947] The title compound was prepared according to Example 5, using 5-bromo-6-methylisoquinoline as the starting material for step 2. MSm / z (M+H + 420.3.

[0948] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(4-methylisoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 123):

[0949]

[0950] Compound 123

[0951] The title compound was prepared according to Example 5, using 5-bromo-4-methylisoquinoline as the starting material for step 2.

[0952]

[0953] Synthesis of 2-([1,2,4]triazolo[4,3-a]pyridin-8-yl)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 124):

[0954]

[0955] Compound 124

[0956] The title compound was prepared according to Example 5, using 8-bromo-[1,2,4]triazolo[4,3-a]pyridine as the starting material for step 2.

[0957]

[0958] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(5,6-dimethoxypyridin-2-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 125):

[0959]

[0960] Compound 125

[0961] The title compound was prepared according to Example 5, using 6-iodo-2,3-dimethoxypyridine as the starting material for step 2.

[0962]

[0963] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1H-indazol-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 126):

[0964]

[0965] Compound 126

[0966] The title compound was prepared according to Example 5, using 4-iodo-1H-indazole as the starting material for step 2.

[0967]

[0968] Synthesis of 3-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)benzonitrile (compound 130):

[0969]

[0970] Compound 130

[0971] The title compound was prepared according to Example 5, using 3-bromobenzonitrile as the starting material for step 2.

[0972]

[0973] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 138):

[0974]

[0975] Compound 138

[0976] The title compound was prepared according to Example 5, using 4-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole as the starting material for step 2.

[0977]

[0978] Synthesis of 6-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)isoindoline-1-one (compound 139):

[0979]

[0980] Compound 139

[0981] The title compound was prepared according to Example 5, using 6-bromoisoindoline-1-one as the starting material for step 2.

[0982]

[0983] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(phthalazin-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 140):

[0984]

[0985] Compound 140

[0986] The title compound was prepared according to Example 5, using 5-bromophthalazine as the starting material for step 2.

[0987]

[0988] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1-(ethoxymethoxy)isoquinoline-7-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 146):

[0989]

[0990] Compound 146

[0991] The title compound was prepared according to Example 5, using 7-bromo-1-(ethoxymethoxy)isoquinoline as the starting material for step 2.

[0992]

[0993] Example 5a

[0994] The following process describes the synthesis of exemplary compounds.

[0995]

[0996] HCl (6N, 0.168 ml, 1.010 mmol) was added to a solution of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2-methoxyquinoline-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (0.22 g, 0.505 mmol, prepared according to Example 5 using 8-bromo-2-methoxyquinoline as the starting material for step 2) in THF (2 ml). The mixture was stirred overnight at 60 °C. The reaction mixture was cooled to room temperature, quenched with 4N NaOH to pH=10, and extracted with EtOAc (twice). The combined organic layers were washed with brine, dried over MgSO4, and concentrated. The crude product was purified by ISCO to give 8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 119).

[0997]

[0998] Synthesis of 6-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-methoxypyridine-2(1H)-one (compound 127):

[0999]

[1000] Compound 127

[1001] The title compound was prepared according to Example 5a, using 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(5,6-dimethoxypyridin-2-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material. MS m / z (M+H + )402.3.

[1002] Synthesis of 7-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)isoquinoline-1(2H)-one (compound 148):

[1003]

[1004] Compound 148

[1005] The title compound was prepared according to Example 5a, using 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1-(ethoxymethoxy)isoquinoline-7-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material.

[1006]

[1007] Synthesis of 8-(5-oxo-3-(piperidin-3-yl)-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 235):

[1008]

[1009] Compound 235

[1010] The title compound was prepared according to Example 5a, using 2-(2-methoxyquinolin-8-yl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material. MS m / z (M+H + 312.1.

[1011] Synthesis of 8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-methoxyquinoline-2(1H)-one (compound 257):

[1012]

[1013] Compound 257

[1014] The title compound was prepared according to Example 5a, using 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2,5-dimethoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material.

[1015]

[1016] Synthesis of (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-methoxyquinoline-2(1H)-one (compound 263):

[1017]

[1018] Compound 263

[1019] The title compound was prepared according to Example 5a, using (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2,5-dimethoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material.

[1020]

[1021] Synthesis of (8-(3-(1-(2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 266):

[1022]

[1023] Compound 266

[1024] The title compound was prepared according to Example 5a, using tert-butyl 6-(3-(5-oxo-1-(2-oxo-1,2-dihydroquinolin-8-yl)-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-2-azaspiro[3.3]heptane-2-carboxylate as the starting material.

[1025]

[1026] Example 5b

[1027] The following process describes the synthesis of exemplary compounds.

[1028]

[1029] TFA (0.402 ml, 5.22 mmol) was added to a solution of 5-((S)-1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (0.25 g, 0.522 mmol, prepared according to Example 5 using 4-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole as the starting material for step 2) in DCM (4 ml). The mixture was stirred overnight at room temperature. LCMS showed only 50% product. Additional TFA (0.402 ml, 5.22 mmol) was added to the mixture. The reaction mixture was stirred overnight at room temperature. LCMS showed 22% of the starting material remaining. Additional TFA (0.402 mL, 5.22 mmol) was added to the mixture, and the reaction mixture was stirred overnight at room temperature. The solvent was removed. The crude product was purified by ISCO (0–20%, MeOH / DCM) followed by reverse-phase ISCO (0–100% ACN / H₂O, 0.1% TFA) to give (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1H-indazol-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 149).

[1030]

[1031] Synthesis of (S)-5-(1-(6-aminospiro[3.3]heptane-2-yl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 261):

[1032]

[1033] Compound 261

[1034] The title compound was prepared according to Example 5b, using (S)-(6-(3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)spiro[3.3]heptane-2-yl)carbamate as the starting material. MS m / z (M+H + 405.3.

[1035] Synthesis of (S)-5-(1-(2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 262):

[1036]

[1037] Compound 262

[1038] The title compound was prepared according to Example 5b, using (S)-6-(3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester as the starting material.

[1039]

[1040] Synthesis of 8-(3-(1-(6-aminospiro[3.3]heptane-2-yl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 268):

[1041]

[1042] Compound 268

[1043] The title compound was prepared according to Example 5b, using (6-(3-(5-oxo-1-(2-oxo-1,2-dihydroquinolin-8-yl)-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)spiro[3.3]heptane-2-yl)carbamate tert-butyl ester as the starting material.

[1044]

[1045] Example 6

[1046] The following process describes the synthesis of exemplary compounds.

[1047]

[1048] Step 1: A mixture of tert-butyl 3-(5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-carboxylate (1.34 g, 4.99 mmol), 8-bromo-2-methoxyquinoline (1.308 g, 5.49 mmol), potassium phosphate (2.120 g, 9.99 mmol), and copper iodide (I) (0.380 g, 1.998 mmol) in DMF (25 mL) was purged with N2 for 5 min. Then (1R,2R)-N,N'-dimethylcyclohexane-1,2-diamine (0.630 mL, 4.00 mmol) was added. The dark brown mixture was stirred at 100 °C for 22 h. LC / MS showed only the starting material. Add another portion of copper iodide (I) (0.380 g, 1.998 mmol), followed by 8-bromo-2-methoxyquinoline (600 mg). The mixture was purged with N2 for 5 min and treated with (1R,2R)-N,N'-dimethylcyclohexane-1,2-diamine (0.630 mL, 4.00 mmol). The reaction mixture was stirred at 100 °C for 24 h. LC / MS showed the formation of new peaks of the desired quality, along with some of the starting material. The reaction was quenched with water and extracted with EtOAc. The combined organic layers were washed with water and concentrated. The solvent was removed, and the residue was purified by rapid chromatography (50 g column, 25 mL / min flow rate, gradient elution: from hexane to EtOAc in 10 min, hold in EtOAc for 5 min, then gradient elution to 15% MeOH / CH2Cl2 in 10 min, then hold in 15% MeOH / CH2Cl2 for 10 min). The fractions containing the desired product were combined (eluted in EtOAc and 5% MeOH / CH2Cl2) and concentrated to give tert-butyl 3-(1-(2-methoxyquinoline-8-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylate (compound 237, 1.2 g, 2.82 mmol, yield 56.5%) as a brown solid.

[1049]

[1050] Step 2: TFA (0.5 mL, 6.49 mmol) was added to a solution of 3-(1-(2-methoxyquinoline-8-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-carboxylic acid tert-butyl ester (compound 237, 0.23 g, 0.541 mmol) in DCM (5 mL). The reaction mixture was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The solvent was removed. The crude product 2-(2-methoxyquinoline-8-yl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 236) was used directly for the next step without further purification. MS m / z (M+H + 326.2.

[1051] Step 3: Potassium carbonate (106 mg, 0.768 mmol) and 1-(2-bromoethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 236, 50 mg, 0.154 mmol) were added to a solution of 2-(2-methoxyquinoline-8-yl)-5-(piperidin-3-yl)-2,4-dichlorobenzene (27.1 µl, 0.169 mmol) in DMF (1.5 mL). The reaction mixture was stirred at 50 °C for 48 hours. The reaction mixture was cooled to room temperature, quenched with water, and extracted with EtOAc (3 times). The combined organic layers were dried over MgSO4 and concentrated. The crude product, 5-(1-(2,4-dichlorophenylethyl)piperidin-3-yl)-2-(2-methoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, was used directly without further purification. MS m / z (M+H) + 498.2.

[1052] Step 4: HCl (6N, 38.8 µl, 0.233 mmol) was added to a solution of 5-(1-(2,4-dichlorophenylethyl)piperidin-3-yl)-2-(2-methoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (29 mg, 0.058 mmol) in THF (1 mL). The mixture was stirred at 60 °C for 5 hours. The reaction mixture was cooled to room temperature, and the solvent was removed. The crude product was purified to give 8-(3-(1-(2,4-dichlorophenylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinolin-2(1H)-one, TFA (compound 241).

[1053]

[1054] Synthesis of 8-(3-(1-(isoindoline-4-ylmethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 238):

[1055]

[1056] Compound 238

[1057] The title compound was prepared according to Example 6, using tert-butyl 4-(bromomethyl)isoindoline-2-carboxylate as the starting material in step 3.

[1058]

[1059] Synthesis of 8-(3-(1-(2,4-dichlorobenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 239):

[1060]

[1061] Compound 239

[1062] The title compound was prepared according to Example 6, using 1-(bromomethyl)-2,4-dichlorobenzene as the starting material in step 3.

[1063]

[1064] Synthesis of 8-(3-(1-(2-chlorobenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 240):

[1065]

[1066] Compound 240

[1067] The title compound was prepared according to Example 6, using 1-(bromomethyl)-2-chlorobenzene as the starting material in step 3.

[1068]

[1069] Synthesis of 8-(5-oxo-3-(1-(2-(pyridin-2-yl)ethyl)piperidin-3-yl)-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 242):

[1070]

[1071] Compound 242

[1072] The title compound was prepared according to Example 6, using 2-(2-bromoethyl)pyridine as the starting material in step 3.

[1073]

[1074] Synthesis of 8-(3-(1-(2,4-dimethylbenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 243):

[1075]

[1076] Compound 243

[1077] The title compound was prepared according to Example 6, using 1-(bromomethyl)-2,4-dimethylbenzene as the starting material in step 3.

[1078]

[1079] Synthesis of 8-(3-(1-((6-methylpyridin-2-yl)methyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 244):

[1080]

[1081] Compound 244

[1082] The title compound was prepared according to Example 6, using 2-(bromomethyl)-6-methylpyridine as the starting material in step 3.

[1083]

[1084] Synthesis of 8-(3-(1-(2-fluoro-4-methoxybenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 245):

[1085]

[1086] Compound 245

[1087] The title compound was prepared according to Example 6, using 1-(bromomethyl)-2-fluoro-4-methoxybenzene as the starting material in step 3.

[1088]

[1089] Synthesis of 8-(5-oxo-3-(1-((1,2,3,4-tetrahydroisoquinoline-8-yl)methyl)piperidin-3-yl)-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 246):

[1090]

[1091] Compound 246

[1092] The title compound was prepared according to Example 6, using tert-butyl 8-(bromomethyl)-1,2,3,4-tetrahydroisoquinoline-2(1H)-formate as the starting material in step 3.

[1093]

[1094] Synthesis of 8-(3-(1-(2-fluorobenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 248):

[1095]

[1096] Compound 248

[1097] The title compound was prepared according to Example 6, using 1-(bromomethyl)-2-fluorobenzene as the starting material in step 3.

[1098]

[1099] Synthesis of 8-(3-(1-(3-methylbenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 249):

[1100]

[1101] Compound 249

[1102] The title compound was prepared according to Example 6, using 1-(bromomethyl)-3-methylbenzene as the starting material in step 3.

[1103]

[1104] Synthesis of 8-(3-(1-(3,5-dimethylbenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 250):

[1105]

[1106] Compound 250

[1107] The title compound was prepared according to Example 6, using 1-(bromomethyl)-3,5-dimethylbenzene as the starting material in step 3.

[1108]

[1109] Synthesis of 8-(3-(1-((4,6-dimethylpyridin-2-yl)methyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 251):

[1110]

[1111] Compound 251

[1112] The title compound was prepared according to Example 6, using 2-(bromomethyl)-4,6-dimethylpyridine as the starting material in step 3.

[1113]

[1114] Synthesis of 8-(3-(1-(4-chloro-2-fluorobenzyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 253):

[1115]

[1116] Compound 253

[1117] The title compound was prepared according to Example 6, using 1-(bromomethyl)-4-chloro-2-fluorobenzene as the starting material in step 3.

[1118]

[1119] Synthesis of 8-(3-(1-(2-(4-chloropyridin-2-yl)ethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 254):

[1120]

[1121] Compound 254

[1122] The title compound was prepared according to Example 6, using 2-(2-bromoethyl)-4-chloropyridine as the starting material in step 3.

[1123]

[1124] Example 6a

[1125] The following process describes the synthesis of exemplary compounds.

[1126]

[1127] Step 1: To a solution of 2-(2-methoxyquinoline-8-yl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (50 mg, 0.154 mmol, prepared according to Example 6, Step 3) in 1,2-dichloroethane (1.5 mL), DIPEA (81 µl, 0.461 mmol), acetic acid (13.20 µl, 0.231 mmol), and 2-(naphthyl-2-yl)acetaldehyde (26.2 mg, 0.154 mmol) were added. The mixture was stirred at room temperature for 10 minutes, and then sodium triacetoxyborohydride (65.1 mg, 0.307 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with water and extracted with DCM (3 times). The combined organic layers were dried and concentrated to give 2-(2-methoxyquinolin-8-yl)-5-(1-(2-(naphthyl-2-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one. MS m / z (M+H + 480.2.

[1128] Step 2: HCl (6N, 34.8 µl, 0.209 mmol) was added to a solution of 2-(2-methoxyquinoline-8-yl)-5-(1-(2-(naphthyl-2-yl)ethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (25 mg, 0.052 mmol) in THF (1 mL). The mixture was stirred at 60 °C for 3 hours. The reaction mixture was cooled to room temperature and the solvent was removed. The crude product was submitted for purification to give 8-(3-(1-(2-(naphthyl-2-yl)ethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 247).

[1129]

[1130] Synthesis of 8-(3-(1-(2-(naphthyl-1-yl)ethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 252):

[1131]

[1132] Compound 252

[1133] The title compound was prepared according to Example 6a, using 2-(naphth-1-yl)acetaldehyde as the starting material in step 1.

[1134]

[1135] Synthesis of (S)-5-(1-(2-(4,4-difluorocyclohexyl)ethyl)piperidin-3-yl)-2-(2-methoxyquinoline-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one, TFA (compound 299):

[1136]

[1137] Compound 299

[1138] The title compound was prepared according to Example 6a, using 2-(4,4-difluorocyclohexyl)acetaldehyde as the starting material in step 1.

[1139]

[1140] Synthesis of (S)-8-(5-oxo-3-(1-(2-(piperidin-4-yl)ethyl)piperidin-3-yl)-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, 2TFA (compound 300):

[1141]

[1142] Compound 300

[1143] The title compound was prepared according to Example 6a, using tert-butyl 4-(2-oxoethyl)piperidine-1-carboxylate as the starting material in step 1.

[1144]

[1145] Synthesis of (S)-8-(3-(1-(2-(4,4-difluorocyclohexyl)ethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 301):

[1146]

[1147] Compound 301

[1148] The title compound was prepared according to Example 6a, using 2-(4,4-difluorocyclohexyl)acetaldehyde as the starting material in step 1.

[1149]

[1150] Synthesis of (S)-8-(5-oxo-3-(1-(2-(tetrahydro-2H-pyran-4-yl)ethyl)piperidin-3-yl)-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, TFA (compound 302):

[1151]

[1152] Compound 302

[1153] The title compound was prepared according to Example 6a, using 2-(tetrahydro-2H-pyran-4-yl)acetaldehyde as the starting material in step 1.

[1154]

[1155] Example 6b

[1156] The following process describes the synthesis of exemplary compounds.

[1157]

[1158] Step 1: A mixture of tert-butyl 3-(5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-carboxylate (1.34 g, 4.99 mmol), 5-bromoisoquinoline (1.308 g, 5.49 mmol), potassium phosphate (2.120 g, 9.99 mmol), and copper iodide (I) (0.380 g, 1.998 mmol) in DMF (25 mL) was purged with N2 for 5 min. Then, trans-N,N'-dimethylcyclohexane-1,2-diamine (0.630 mL, 4.00 mmol) was added. The dark brown mixture was stirred at 100 °C for 22 h. Another portion of copper iodide (I) (0.380 g, 1.998 mmol) was added, followed by 5-bromoisoquinoline (600 mg). The mixture was purged with N2 for 5 min and treated with trans-N,N'-dimethylcyclohexane-1,2-diamine (0.630 mL, 4.00 mmol). The reaction mixture was stirred at 100 °C for 24 h. The reaction was quenched with water and extracted with EtOAc. The combined organic layers were washed with water and concentrated. The solvent was removed, and the residue was purified by rapid chromatography. The fractions containing the desired product were combined and concentrated to give tert-butyl 3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidine-1-carboxylate (1.2 g, 2.82 mmol, 56.5% yield) as a brown solid.

[1159] Step 2: Add TFA (0.5 mL, 6.49 mmol) to a solution of tert-butyl piperidine-1-carboxylate (0.23 g, 0.541 mmol) in DCM (5 mL). Stir the reaction mixture overnight at room temperature. Remove the solvent. The crude product is used directly in the next step without further purification. MS m / z (M+H + 326.2.

[1160] Step 3: Hunig base (271 µl, 1.549 mmol) and sodium triacetoxyborohydride (197 mg, 0.929 mmol) were added to a mixture of (S)-2-(isoquinolin-5-yl)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (183 mg, 0.620 mmol) and 6-oxo-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (196 mg, 0.929 mmol) in THF (6 mL). The slurry reaction mixture was stirred for 1 hour. The reaction was quenched with 1 N NaOH solution (0.2 mL). The mixture was concentrated and purified by rapid chromatography. The fractions eluted in 15% MeOH / CH2Cl2 containing the desired product were combined and concentrated to give (S)-6-(3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (compound 258, 230 mg, 0.469 mmol, yield 76%).

[1161]

[1162] Synthesis of 6-((S)-3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-2-azaspiro[3,4]octane-2-carboxylic acid tert-butyl ester (compound 255):

[1163]

[1164] Compound 255

[1165] The title compound was prepared according to Example 6b, using tert-butyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate as the starting material in step 3.

[1166]

[1167] Synthesis of (6-((S)-3-(1-(isoquinolin-5-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)spiro[3.3]heptane-2-yl)tert-butyl carbamate (compound 259):

[1168]

[1169] Compound 259

[1170] The title compound was prepared according to Example 6b, using (6-oxospiro[3.3]heptane-2-yl)carbamate tert-butyl ester as the starting material in step 3.

[1171]

[1172] Synthesis of (5-(1-(2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-2-(2-methoxyquinoline-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 264):

[1173]

[1174] Compound 264

[1175] The title compound was prepared according to Example 5b, using tert-butyl 6-(3-(1-(2-methoxyquinoline-8-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-2-azaspiro[3.3]heptane-2-carboxylate as the starting material, which itself was prepared according to Example 6b using 8-bromo-2-methoxyquinoline as the starting material for step 1.

[1176]

[1177] Example 7

[1178] The following process describes the synthesis of exemplary compounds.

[1179]

[1180] Step 1: Add DIPEA (1.280 mL, 7.33 mmol), sodium triacetoxyborohydride (1.553 g, 7.33 mmol), and 2-cyclohexylacetaldehyde (0.805 mL, 5.86 mmol) to a suspension of (S)-5-(piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one hydrochloride (1 g, 4.89 mmol) in THF (25 mL). Stir the mixture at room temperature for 2 hours. Quench the reaction with water. Filter the solid and dry it. Extract the filtrate with DCM (3 times). Dry and concentrate the combined organic layers. The crude product (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one) was used directly for the next step without further purification. MS m / z (M+H + 279.3.

[1181] Step 2: Under N2, 8-bromo-2-methoxyquinoline (1.668 g, 7.00 mmol) was added to a suspension of (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (1.3 g, 4.67 mmol), copper iodide (I) (0.356 g, 1.868 mmol), and potassium carbonate (3.23 g, 23.35 mmol) in DMF (40 ml), followed by trans-N,N'-dimethylcyclohexane-1,2-diamine (0.589 ml, 3.74 mmol). The reaction mixture was stirred at 100 °C for 2 days. LC-MS showed that a significant amount of starting material remained. Copper iodide (I) (0.356 g, 1.868 mmol) and trans-N,N'-dimethylcyclohexane-1,2-diamine (0.589 mL, 3.74 mmol) were added to the reaction mixture. The reaction mixture was stirred at 100 °C for 3 days. The reaction mixture was cooled to room temperature and quenched with water. The solid was filtered and washed with DCM. The filtrate was concentrated. The crude product was purified by ISCO (0–20%, MeOH / EtOAc) to give (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2-methoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (0.75 g, 36.9%). MS m / z (M+H + )436.3.

[1182] Step 3. Add HCl (6N, 1.224 mL, 7.35 mmol) to a solution of (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2-methoxyquinoline-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (0.8 g, 1.837 mmol) in THF (16 mL). Stir the mixture overnight at 60 °C. Cool the reaction mixture to room temperature and remove the solvent. Add water to the residue and extract with DCM (6 times). Dry the combined organic layers with MgSO4 and concentrate. The crude product was purified by ISCO (0-20%, MeOH / DCM) to give (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, HCl (compound 144).

[1183]

[1184] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 28):

[1185]

[1186] Compound 28

[1187] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 1-iodo-3-methoxybenzene as the starting material in step 2. MS m / z (M+H) + 371.3.

[1188] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(2-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 29):

[1189]

[1190] Compound 29

[1191] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 1-iodo-2-methoxybenzene as the starting material in step 2.

[1192]

[1193] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(isoquinolin-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 62):

[1194]

[1195] Compound 62

[1196] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 5-iodoisoquinoline as the starting material in step 2.

[1197]

[1198] Synthesis of 5-(3-(1-(cyclohexylmethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3,4-dihydroisoquinoline-1(2H)-one (compound 72):

[1199]

[1200] Compound 72

[1201] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and tert-butyl 5-bromo-1-oxo-3,4-dihydroisoquinoline-2(1H)-formate as the starting material in step 2.

[1202]

[1203] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(2,3-dihydrobenzo[b][1,4]dioxane-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 73):

[1204]

[1205] Compound 73

[1206] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 6-iodo-2,3-dihydrobenzo[b][1,4]dioxane as the starting material in step 2.

[1207]

[1208] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(benzo[d][1,3]dioxacyclopenten-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 74):

[1209]

[1210] Compound 74

[1211] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 5-bromobenzo[d][1,3]dioxane as the starting material in step 2.

[1212]

[1213] Synthesis of tert-butyl 5-(3-(1-(cyclohexylmethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-1-oxo-3,4-dihydroisoquinoline-2(1H)-formate (compound 80):

[1214]

[1215] Compound 80

[1216] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and tert-butyl 5-bromo-1-oxo-3,4-dihydroisoquinoline-2(1H)-formate as the starting material in step 2.

[1217]

[1218] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(quinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 84):

[1219]

[1220] Compound 84

[1221] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 8-iodoquinoline as the starting material in step 2.

[1222]

[1223] Synthesis of 2-(benzo[d][1,3]dioxacyclopenten-4-yl)-5-(1-(cyclohexylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 87):

[1224]

[1225] Compound 87

[1226] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 4-iodobenzo[d][1,3]dioxane as the starting material in step 2.

[1227]

[1228] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(quinolin-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 89):

[1229]

[1230] Compound 89

[1231] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 5-iodoquinoline as the starting material in step 2.

[1232]

[1233] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(isoquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 99):

[1234]

[1235] Compound 99

[1236] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 8-iodoisoquinoline as the starting material in step 2.

[1237]

[1238] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(1H-indazol-7-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 103):

[1239]

[1240] Compound 103

[1241] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 7-bromo-1H-indazole as the starting material in step 2.

[1242]

[1243] Synthesis of 8-(3-(1-(cyclohexylmethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 109):

[1244]

[1245] Compound 109

[1246] The title compound was prepared according to Example 5a, using 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(2-methoxyquinoline-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material. It itself was prepared according to Example 7 using cyclohexaneformaldehyde as the starting material in step 1 and 8-bromo-2-methoxyquinoline as the starting material in step 2.

[1247]

[1248] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-4-yl)-2-(isoquinolin-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 112):

[1249]

[1250] Compound 112

[1251] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 5-iodoisoquinoline as the starting material in step 2.

[1252]

[1253] Synthesis of 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(2-methoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 113):

[1254]

[1255] Compound 113

[1256] The title compound was prepared according to Example 7, using cyclohexaneformaldehyde as the starting material in step 1 and 8-bromo-2-methoxyquinoline as the starting material in step 2.

[1257]

[1258] Synthesis of 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(2,5-dimethoxyquinolin-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 256):

[1259]

[1260] Compound 256

[1261] The title compound was prepared according to Example 7, using 8-bromo-2,5-dimethoxyquinoline as the starting material in step 2.

[1262]

[1263] Example 7a

[1264] The following process describes the synthesis of exemplary compounds.

[1265]

[1266] At 0 °C, BBr3 (1.329 ml, 1.329 mmol, 1 M solution in DCM) was added dropwise to a solution of (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-methoxyquinoline-2(1H)-one (150 mg, 0.332 mmol) in DCM (3 ml). The reaction mixture was stirred at room temperature for 5 days. After the reaction was complete, water (10 ml) was added to the reaction mixture at 0 °C. The mixture was extracted with EtOAc (20 ml x 3). The combined organic layers were washed with saturated NaHCO3. The aqueous layer was concentrated to give (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-hydroxyquinoline-2(1H)-one (compound 296).

[1267]

[1268] Example 7b

[1269] The following process describes the synthesis of exemplary compounds.

[1270]

[1271] Step 1: Hunig base (8.83 mL, 50.6 mmol), cyclohexaneformaldehyde (2.94 mL, 24.27 mmol), and sodium triacetoxyborohydride (6.43 g, 30.3 mmol) were added to a mixture of 5-(piperidin-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one hydrochloride (4.14 g, 20.23 mmol) in THF (50 mL). The reaction mixture was stirred for 1.5 hours. The reaction was quenched with 1 N NaOH solution (5 mL). The mixture was concentrated and purified by rapid chromatography. The fractions eluted from 20% to 25% MeOH / CH2Cl2 were combined and concentrated to give 5-(1-(cyclohexylmethyl)piperidin-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (3.42 g, 9.04 mmol, yield 44.7%) as a grayish-white solid.

[1272] Step 2: A mixture of 5-(1-(cyclohexylmethyl)piperidin-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (65 mg, 0.246 mmol), 1-iodo-4-methoxybenzene (86 mg, 0.369 mmol), potassium carbonate (104 mg, 0.492 mmol), and copper iodide (I) (18.73 mg, 0.098 mmol) in DMF (1 mL) was purged with N2 for 3 min. Then (1R,2R)-N,N'-dimethylcyclohexane-1,2-diamine (31.0 µl, 0.197 mmol) was added. The blue mixture was stirred at 100 °C for 44 h. The reaction was quenched with 1 N NaOH solution. The mixture was extracted with EtOAc, washed with water and brine, and concentrated. The residue was purified by rapid chromatography to give 5-(1-(cyclohexylmethyl)piperidin-4-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 111).

[1273]

[1274] Example 8

[1275] The following process describes the synthesis of exemplary compounds.

[1276]

[1277] Step 1: Add NaH (9.32 mg, 0.233 mmol) to a solution of (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-hydroxyquinoline-2(1H)-one (34 mg, 0.078 mmol) in DMF (1 mL). Stir the mixture at room temperature for 30 minutes. Add tert-butyl (3-bromopropyl)carbamate (20.35 mg, 0.085 mmol) to the reaction mixture. Stir the reaction mixture at room temperature overnight. Add water to the mixture and extract with EtOAc (twice). Wash the combined organic layers with brine, dry over MgSO4, and concentrate. Use the crude product directly for the next step without further purification. MS m / z (M+H + 595.4.

[1278] Step 2: To a solution of (S)-(3-((8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-oxo-1,2-dihydroquinoline-5-yl)oxy)propyl)carbamate tert-butyl ester (20 mg, 0.034 mmol) in DCM (1 ml), TFA (0.1 ml, 1.298 mmol) was added. The reaction mixture was stirred overnight at room temperature. The solvent was removed. The crude product was submitted for purification to give (S)-5-(3-aminopropoxy)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one, 2 TFA (compound 298).

[1279]

[1280] Example 8a

[1281] The following process describes the synthesis of exemplary compounds.

[1282]

[1283] To a solution of (S)-5-(3-aminopropoxy)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 298, 32 mg, 0.065 mmol) in DMF (0.6 mL), 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (21.44 mg, 0.078 mmol) and DIPEA (56.5 µl, 0.323 mmol) were added. The reaction mixture was stirred overnight at 80 °C. The reaction mixture was cooled, and then water was added to the mixture. The resulting solid was filtered and washed with water. The crude product was dissolved in DMF and purified to give 4-((3-((8-(3-((S)-1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-oxo-1,2-dihydroquinoline-5-yl)oxy)propyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione, TFA (compound 297). MS m / z (M+H + 751.1.

[1284] Example 9

[1285] The following process describes the synthesis of exemplary compounds.

[1286]

[1287] To a solution of 2-(4-methoxyphenyl)-5-((3S)-1-(piperidin-3-ylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (65 mg, 0.175 mmol) in DCM (2 mL), add 46.9 µl, 0.350 mmol of trimethyl isocyanate. Stir the reaction mixture at room temperature for 3 hours. Add another portion of trimethyl isocyanate (46.9 µl, 0.350 mmol). Stir the reaction mixture at room temperature for 2 days. The solvent was evaporated, and the crude product was purified by ISCO (0-20% MeOH / EtOAc) to give 2-(4-methoxyphenyl)-5-((3S)-1-(1-formamidopiperidin-3-ylmethyl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 91).

[1288]

[1289] Example 10

[1290] The following process describes the synthesis of exemplary compounds.

[1291]

[1292] To a solution of 8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (18 mg, 0.043 mmol) in EtOH (2 mL) / H₂O (0.1 mL), 10 wt% Pd / C (5 mg) and ammonium formate (26.9 mg, 0.427 mmol) were added. The reaction mixture was stirred at 80 °C for 15 hours. The mixture was filtered through a syringe filter and washed with MeOH. The filtrate was concentrated and purified by rapid chromatography to give 8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3,4-dihydroquinoline-2(1H)-one (compound 129).

[1293]

[1294] Example 11

[1295] The following process describes the synthesis of exemplary compounds.

[1296]

[1297] Concentrated HCl solution was added to a vial containing 30 mg (0.079 mmol) of 3-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)benzonitrile. The brown mixture was stirred at room temperature for 28 hours. The solvent was removed, and the residue was purified by rapid chromatography to give 3-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)benzamide (compound 132).

[1298]

[1299] Example 12

[1300] The following process describes the synthesis of exemplary compounds.

[1301]

[1302] Hunig base (161 µl, 0.922 mmol) was added to a mixture of (S)-5-(1-(2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-2-(isoquinoline-5-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (90 mg, 0.230 mmol) in CH2Cl2 (2 mL) / THF (2 mL). After stirring for 3 minutes, a suspension was formed. Then paraformaldehyde (69.2 mg, 2.305 mmol) and sodium triacetoxyborohydride (147 mg, 0.691 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was quenched with a few drops of 1N NaOH solution, concentrated, and purified by reversed-phase rapid chromatography (5g C18 gold column, gradient elution: from water containing 0.1% NH4OH to 40% CH3CN / H2O containing 0.1% NH4OH) to give (S)-2-(isoquinolin-5-yl)-5-(1-(2-methyl-2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 260, 8 mg, 0.020 mmol, yield 8.58%) as a white powder. MS m / z (M+H + 405.2.

[1303] Synthesis of 8-(3-(1-(2-methyl-2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 265):

[1304]

[1305] Compound 265

[1306] The title compound was prepared according to Example 12, using 8-(3-(1-(2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one as the starting material. MS m / z (M+H + 421.3.

[1307] Synthesis of 2-(2-methoxyquinoline-8-yl)-5-(1-(2-methyl-2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 267):

[1308]

[1309] Compound 267

[1310] The title compound was prepared according to Example 12, using 5-(1-(2-azaspiro[3.3]heptane-6-yl)piperidin-3-yl)-2-(2-methoxyquinoline-8-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one as the starting material.

[1311]

[1312] Example 13 – Chiral separation of inhomogeneities

[1313] Preparation of (S)-5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 4) and (R)-5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 5):

[1314]

[1315] Compound 4 Compound 5

[1316] The title compound was prepared by chiral HPLC separation of the racemic mixture 5-(1-(cyclohexylmethyl)piperidin-3-yl)-2-(4-methoxyphenyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (33 mg) using a ChiralCel® OJ (5 x 50 cm, 20 µm) with methanol / diethylamine 100:0.1 as the mobile phase. Compound 4: Optical rotation – positive; ee 97.7%; MS m / z (M+H)+ 371.5. Compound 5: Optical rotation – negative; ee > 98%. MS m / z (M+H + 371.5.

[1317] Preparation of (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3,4-dihydroquinoline-2(1H)-one (compound 142) and (R)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3,4-dihydroquinoline-2(1H)-one (compound 143):

[1318]

[1319] Compound 142 Compound 143

[1320] The title compound was prepared by chiral HPLC separation of the racemic mixture 8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3,4-dihydroquinoline-2(1H)-one (33 mg) using a ChiralPak® IG (5 x 50 cm, 20 µm) with methanol / diethylamine 100:0.1 as the mobile phase. Compound 142: Optical rotation – positive; ee 97.7%; MS m / z (M+H) + 424.3. Compound 143: Optical rotation – negative; ee 97.7%. MS m / z (M+H) + 424.3.

[1321] Preparation of (S)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 144) and (R)-8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (compound 145):

[1322]

[1323] Compound 144 Compound 145

[1324] The title compound was prepared by chiral HPLC separation of the racemic mixture 8-(3-(1-(2-cyclohexylethyl)piperidin-3-yl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)quinoline-2(1H)-one (33 mg) using a ChiralPak® IG (5 x 50 cm, 20 µm) with methanol / diethylamine 100:0.1 as the mobile phase. Compound 144: Optical rotation – positive; ee 97.7%; MS m / z (M+H) + 422.3. Compound 145: Optical rotation – negative; ee 97.7%. MS m / z (M+H) + )422.3.

[1325] Preparation of (S)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1H-indazol-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 149) and (R)-5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1H-indazol-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 150):

[1326]

[1327] Compound 149 Compound 150

[1328] The title compound was prepared by chiral HPLC separation of the racemic mixture 5-(1-(2-cyclohexylethyl)piperidin-3-yl)-2-(1H-indazol-4-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (33 mg) using a ChiralPak® IG (5 x 50 cm, 20 µm) with a mobile phase of hexane / EtOH / iPrNH2 40:60:0.04. Compound 149: Optical rotation – positive; ee 97.7%; MS m / z (M+H) + 395.2. Compound 150: Optical rotation – negative; ee 97.7%; MS m / z (M+H) + )395.2.

[1329] Table 2 shows other compounds synthesized using a similar procedure as described herein.

[1330] Table 2

[1331]

[1332]

[1333]

[1334]

[1335]

[1336]

[1337]

[1338]

[1339]

[1340]

[1341]

[1342]

[1343] Biological research

[1344] Example 14. High-throughput screening (HTS) NNMT inhibition assay

[1345] Determination of measurement parameters

[1346] The initial rate method was used to determine the K-value of nicotinamide for nicotinamide N-methyltransferase (NNMT). M Value. In short, MTase enzyme was diluted to 5 to 30 nM in 20 mM Tris (pH 8.0), 150 mM NaCl, 12 mM MgCl2, 4 mM EDTA, 0.1% BSA, and 100 μM S-adenosyl-L-methionine (SAM). The reaction was initiated by adding different concentrations of substrate. The reaction was carried out at appropriate time points, and MTase-Glo was added... TM The reagents (Promega, Madison, Wisconsin) terminated the reaction process by consuming excess SAM. MTase-Glo was used. TM The kit (Promega) measures reaction progress by the degree to which SAM is converted to S-adenosine-L-homocysteine ​​(SAH). The K+ of SAM is determined using the same method. M The concentration of nicotinamide was kept constant at 100 μM, while the concentration of SAM was varied. The initial rate was determined by plotting SAH generation as a function of time to generate a linear slope. These rates were then plotted as a function of substrate concentration and fitted to the Michaelis-Menten equation to determine K. M value.

[1347] Using previously determined nicotinamide and SAM K M Time-process experiments were conducted to determine the HTS assay duration. Solutions of 5 μM SAM and 5 μM nicotinamide were prepared in 20 mM Tris (pH 8.0), 150 mM NaCl, 12 mM MgCl2, 4 mM EDTA, and 0.1% BSA. The reaction was initiated by adding small molecule methyltransferase (SMMTase) to final concentrations of 7.5 μM, 15 μM, or 30 μM. The reaction was initiated by adding MTase-Glo TM The reagents were used to consume excess SAM, and the reaction progress was recorded periodically. The 20% conversion point was determined by comparing the reaction progress with the SAH standard curve. The ideal reaction time and enzyme concentration were selected based on combinations that achieved 10% to 20% conversion within 20 to 30 minutes.

[1348] The NNMT activity assay for HTS was performed according to the protocol described in Table 3.

[1349] Table 3. NNMT HTS MTase-Glo TM Measurement scheme

[1350]

[1351]

[1352]

[1353] Table 4 shows the IC50 values ​​of the compounds disclosed herein in the NNMT activity HTS assay. 50 In Table 4, the plus sign indicates the following meanings: ++++ = IC 50 ≤ 100 nM; +++ = IC 50 > 100 nM and ≤ 1 µM; ++ = IC 50 > 1 µM and ≤ 10 µM; + = IC 50 > 10 µM; null = no significant inhibition.

[1354] Table 4

[1355]

[1356]

[1357]

[1358] Example 15. Evaluation of compounds 144 and 145 in a mouse tumor model.

[1359] mouse tumor model

[1360] In a high-grade serous ovarian cancer model, two million ID8-luc ovarian cancer cells were intraperitoneally (ip) injected into female C57BL / 6 mice, and body weight was measured weekly. Mice were sacrificed at indicated time points, and the omental tumor weight was measured. In a subcutaneous tumor model, one million MC38 colon cancer cells or five million low-grade serous ovarian cancer cell line LG-6406 cells were subcutaneously (sc) injected into the flanks of C57BL / 6 mice or nude mice, respectively. Tumor volume was measured twice weekly along three axes (length a, width b, and height c), calculated using the formula: Tumor volume = (a × b × c) / 2. The maximum tumor size (2000 mm) was measured. 3 Mice were euthanized before the tumor diameter reached 1.5 cm (as approved by the University of Chicago IACUC). In the intravenous tumor model, 100,000 EO771-LMB breast cancer cells were inoculated into C57BL / 6 mice via retroorbital vein (iv) under continuous isoflurane inhalation anesthesia, and the mice were sacrificed 12 to 16 days later. To analyze the number and size of metastases, lungs were removed, formalin-fixed, paraffin-embedded, and stained with H&E by the University of Chicago Human Tissue Resource Center. One slide was analyzed from each lung. Slides were scanned using an Olympus VS200 whole-slide scanner, and metastases were measured in QuPath (v0.3.0). Metastasis sizes were normalized to the total lung area of ​​the corresponding slide, and the average size of the five largest metastases per mouse was plotted.

[1361] Treatment with compound 144 or 145

[1362] For intraperitoneal treatment, the active NNMT inhibitor (NNMTi) compound 144 or its inferior (NNMTi-D) compound 145 was dissolved in 10% DMSO and mixed with alginate (Millipore Sigma, 180947) in a 1:1 ratio. Each mouse was intraperitoneally injected with 100 μL of a solution containing a final concentration of 10 mg / mL alginate and 4 mg / kg NNMTi or its inferior [Downs et al., J. Cell. Physiol., 152:422-429 (1992)]. Mice were intraperitoneally injected with the solvent (10% DMSO, 10 mg / kg alginate), NNMTi (100 µg / mouse), or its inferior (100 µg / mouse). Initial treatment began 7 days after ID8-luc injection, with treatment administered three times weekly until the end of the study. Tumor burden was assessed using an in vivo imaging system (IVIS) (PerkinElmer IVIS® Spectrum). Mice were injected with 200 μL of fluorescein-PBS solution (15 mg / mL) and anesthetized with isoflurane. Imaging was performed after a 7-minute incubation period using the following settings: automatic exposure, medium pixel binning, and F / stop 1. Living Image was used. ® The software (version 4.7.3, PerkinElmer) quantifies the radioactivity. Figure 1 The results showed that in a mouse model of high-grade serous ovarian cancer, treatment with compound 144 (NNMTi) significantly reduced tumor burden compared with treatment with solvent or inferior compound 145 (NNMTi-D).

[1363] For intratumoral (it) injection into subcutaneous MC38 or LG-6406 tumors, NNNMTi or its inverse was dissolved in 10% DMSO and mixed with alginate (Millipore Sigma, 180947) at a 1:1 ratio. A total volume of 50 µL of the solution containing 4 mg / kg NNNMTi or its inverse, and a final alginate concentration of 10 mg / ml, was injected into the tumor. Treatment with the solvent (10% DMSO, 10 mg / kg alginate), NNNMTi (100 µg / mouse), or its inverse (100 µg / mouse) was initiated 5 days after injection into MC38 cancer cells, every three days until day 20 (for the MC38 model); treatment with LG-6406 cancer cells was initiated 7 days after injection, twice weekly until the end of the study (for the LG-6406 model). Tumor burden was measured twice weekly. Figure 2The results showed that in a mouse model of subcutaneous MC38 tumors, treatment with compound 144 (NNMTi) significantly reduced tumor volume compared to treatment with the solvent or inferior compound 145 (NNMTi-D). Five days after subcutaneous injection of other MC38 colon cancer cells, mice were given NNMTi or inferior compound 144 with a PD-1 inhibitor monoclonal antibody (αPD1, InVivoMAb anti-mouse PD-1, clone RPM1-14, BioXCell, Cat#BE0146; RRID: AB_10949053) or an IgG control antibody (InVivoMAb rat IgG). 2a Combination therapy with isotype control, clone 2A3 (BioXCell catalog number BE0089; RRID: AB_1107769). Combination use of NNMTi with immune checkpoint inhibitors: Figure 3A and Figure 3B The results showed that treatment with both compound 144 (NNMTi) and the PD-1 inhibitor (αPD1) significantly and synergistically reduced tumor volume in a mouse subcutaneous MC38 tumor model. Figure 4 The results showed that in a mouse model of low-grade serous ovarian cancer, treatment with compound 144 (NNMTi) significantly reduced tumor volume compared to treatment with the solvent or inferior compound 145 (NNMTi-D). This result is noteworthy because low-grade serous ovarian cancer is particularly resistant to conventional cancer therapies.

[1364] For the venous tumor model, mice were anesthetized with isoflurane, and 50 µL of solvent, NNMTi (4 mg / kg), or its inferior (4 mg / kg) was dispersed in the throat as previously described [Nielsen et al., J. Vis. Exp., 57672 (2018), doi:10.3791 / 57672]. Treatment via oropharyngeal (op) inhalation resulted in local lung treatment. Treatment with solvent (10% DMSO, 10 mg / kg alginate), NNMTi (100 µg / mouse), or its inferior (100 µg / mouse) was initiated two days prior to cancer cell injection and continued daily until five days after cancer cell injection (prevention study). The lung was removed 13 days later. Figure 5The results showed that in a mouse model of intravenous tumors, treatment with compound 144 (NNMTi) significantly reduced the size of metastatic lesions compared with treatment with the solvent or inferior compound 145 (NNMTi-D). Other mice that received intravenous injection of EO771 breast cancer cells, in combination with a PD-1 inhibitor monoclonal antibody (αPD1, InVivoMAb anti-mouse PD-1, clone RPM1-14, BioXCell, Cat#BE0146; RRID: AB_10949053) or an IgG control antibody (InVivoMAb rat IgG2a isotype control, clone 2A3, BioXCell Cat#BE0089; RRID: AB_1107769), were treated with NNNMTi or inferior compounds via oropharyngeal inhalation until five days after cancer cell injection, and the lungs were resected 16 days later. Figure 6 The results showed that in a mouse intravenous tumor model, treatment with compound 144 (NNMTi) plus an IgG control antibody or a PD-1 inhibitor (αPD1) significantly reduced the size of metastatic lesions.

[1365] Whole-body Nnmt- / - mice showed reduced tumor burden.

[1366] Figure 7A and Figure 7B The study showed that whole-body Nnmt- / - mice exhibited a reduced tumor burden. Figure 7A Nnmt+ / + (n=6) and Nnmt- / - (n=6) mice were injected into the left ovarian cyst and injected with the mouse HGS-2 ovarian cancer cell line. The tumor burden of primary ovarian and omental tumors was assessed after 42 days (Student's t-test). Figure 7B Nnmt+ / + (n=6) and Nnmt- / - (n=6) mice were injected into the left ovarian cyst and injected with the mouse HGS-3 ovarian cancer cell line. The tumor burden of primary ovarian and omental tumors was assessed after 42 days (Student's t-test). Figure 8 The results showed that whole-body Nnmt- / - mice exhibited reduced tumor burden. Nnmt-knockout in the tumor microenvironment reduced tumor burden. Nnmt+ / + wild-type (n=10) and Nnmt- / - knockout mice (n=11) were injected into the mammary fat pads, followed by the injection of mouse EO771-LMB breast cancer cell line, and tumor growth was monitored twice weekly by caliper measurement (one-way ANOVA).

[1367] NNMTi treatment reduces tumor burden

[1368] Figure 9A and Figure 9B The results showed that NNMT inhibition significantly reduced tumor burden and metastasis in an orthotopic mouse model of ovarian cancer. Figure 9AC57Bl6 mice were injected with the HGS-2 ovarian cancer cell line into the left ovarian cyst, and NNMTi (formulation: 10% NMP, 20% PEG400, 70% Solutol) was administered intraperitoneally for 5 weeks starting 4 days after cancer cell injection. Figure 9B C57Bl6 mice were injected with BPNM ovarian cancer cells into the left ovarian cyst, and treated daily for 12 days starting 4 days after cancer cell injection with NNMTi at 25 mg / kg via intraperitoneal injection (formulation: 10% NMP, 20% PEG400, 70% Solutol). BPNM (Trp53− / −R172HBrca1− / −Pten− / −Nf1− / −MycOE genotype) was HR-deficient (S. Iyer Cancer Discovery 2021).

[1369] Figure 10 The results showed that NNMT inhibition significantly reduced tumor burden in an orthotopic mouse model of breast cancer. C57Bl6 mice were injected with the mouse EO771-LMB breast cancer cell line into the mammary fat pad, and starting 4 days after cancer cell injection, mice were treated daily by intraperitoneal injection of NNMTi (25 mg / kg) or a solvent control. (Formulation: 10% NMP, 20% PEG400, 70% Solutol) (One-way ANOVA).

[1370] Combination of NNMTi with radiotherapy

[1371] Figure 11 The results showed that NNMT inhibition improved the therapeutic response to local radiotherapy. C57BL / 6 mice were subcutaneously injected with LLC (Lewis lung cancer) cells and treated daily from day 4 via intraperitoneal injection of either solvent or NNMTi. On day 13, the tumors were locally irradiated with 20 Gy (10 mice per group, two-factor ANOVA).

[1372] Effects of NNMT inhibition on CAF in vivo

[1373] Figure 12A , Figure 12B and Figure 12C This demonstrates the effect of NNMT inhibition on CAF in vivo. Figure 12AEffects of NNMT inhibition on CAF in vivo in a breast cancer model. NNMT inhibition reduced the abundance of established CAF subtypes in vivo. Flow cytometry: C57Bl6 mice were injected with the mouse EO771-LMB breast cancer cell line into the mammary fat pad. Four days after injection, they were treated daily by intraperitoneal injection with NNMTi (25 mg / kg) or a solvent control. Eighteen days after cancer cell injection, the tumors were dissected and analyzed by flow cytometry. Ly6C+ inflammatory CAF (iCAF), α-SMA+ myofibroblast CAF (myCAF), MHCII+ antigen-presenting CAF (apCAF), and PDGFRα+ vascular CAF (vCAF) were identified. Figure 12B In an orthotopic ovarian cancer model, NNMTi treatment reduced the abundance of Ly6C+iCAF and MHCII+myCAF. Flow cytometry (gated): C57Bl6 mice were injected with the mouse HGS-2 ovarian cancer cell line into their ovarian cysts. Four days post-injection, they were treated daily via intraperitoneal injection with either NNMTi (25 mg / kg) or a solvent-based control. Nineteen days post-injection, the tumors were dissected and analyzed by flow cytometry. Figure 12C NNMTi treatment increased H3K27 trimethylation in CAF in vivo. Flow cytometry: Absolute quantification of low H3K27me expression in CD45-EPCAM-CD31-Podoplanin+ CAF in primary EO771-LMB tumors treated with NNMTi or solvent control.

[1374] Effect of NNMT inhibition on complement factor C3 abundance

[1375] Figure 13 The effect of NNMT inhibition on complement factor C3 abundance was shown. NNMTi treatment reduced complement factor secretion in vivo. Enzyme-linked immunosorbent assay (ELISA): EO771-LMB primary tumor homogenates treated with NNMTi or solvent control were homogenized 18 days after cancer cell injection, and complement factor C3 protein was detected (Student's t-test).

[1376] The effect of NNMTi treatment on tumor immune cell infiltration

[1377] Figure 14A , Figure 14B and Figure 14C This study demonstrates the effect of NNMTi treatment on tumor immune cell infiltration. NNMTi treatment reduced the number of immunosuppressive monocytes of myeloid origin (M-MDSCs; Ly6Cs). 高 PD-L1 高 The abundance of ) and increased CD8+ T cell response. Figure 14AMC38 colon cancer cells were subcutaneously (sc) injected into C57BL / 6 mice (day 0). Five days later, once the tumor was palpable, mice were treated every 3 days with intratumoral injection (it) of either NNMTi or NNMTi-D as a control. Fifteen days after cancer cell injection, MC38 tumors were dissected and subjected to 19-color flow cytometry (6-8 mice per group, Student's t-test). Figure 14B Fifteen days after injection of cancer cells, MC38 tumors were dissociated, stimulated with PMA / ionomycin, and the production of effector cytokines was quantified (Student's t-test). Figure 14C C57Bl6 mice were orthotopically injected with HGS-2 ovarian cancer cells or EO771-LMB breast cancer cells. Four days after cancer cell injection, mice were treated daily with NNMTi via intraperitoneal injection at a dose of 25 mg / kg. Eighteen days after cancer cell injection, the primary tumors were dissected and analyzed by flow cytometry (Student's t-test).

[1378] The impact of NNMTi treatment on the tumor metabolome

[1379] Figure 15A and Figure 15B This study demonstrates the effects of NNMTi treatment on the tumor metabolome. NNMTi treatment locally reduced the levels of 1-methylnicotinamide (1-MNA) and its metabolite N1-methyl-2-pyridone-5-carboxamide (2PY) in tumor tissue, while simultaneously leading to the accumulation of S-adenosylmethionine (SAM). Figure 15A Experimental plan: MC38 cells were subcutaneously injected into C57BL / 6 mice. Thirteen days later, the mice were treated with intratumoral injection of the solvent, NNNMTi-D, or NNNMTi. Tumors were then sampled at different time points after injection. Figure 15B Tumor samples were extracted for liquid chromatography-mass spectrometry analysis. Central metabolites of the methionine and nicotinamide cycles were shown to reflect the effects of NNMT inhibition on the levels of these metabolites (n=5 mice / group. Conventional one-way ANOVA).

[1380] NNMT inhibition exerts its tumor-reducing effect by targeting the tumor microenvironment (TME).

[1381] Figure 16This demonstrates that NNMT inhibition exerts its tumor-reducing effect by targeting the TME, rather than by altering cancer cell growth. To determine whether NNMT inhibition in cancer cells reduces tumor growth in vivo, we injected MC38 colon cancer cells into systemically Nnmt- / - mice and administered either a solvent or NNMTi. These mice were completely lacking in NNMT, so NNMTi treatment should have no specific effect on TME-mediated cells. Since the cancer cells were Nnmt-wildtype and expressed NNMT, any effect of NNMT inhibition on cancer cell growth would be detectable. However, no tumor shrinkage was observed in our experiments.

[1382] It should be understood that although specific embodiments have been shown and described herein, various modifications can still be made to the present invention, and all such modifications are within the scope of this disclosure. Furthermore, this disclosure is not limited to the specific embodiments listed herein. The descriptions and illustrations of the embodiments and implementations herein should not be construed as limiting this disclosure. Moreover, the entire content of this disclosure is not limited to the specific illustrations, structures, or relative proportions shown herein; the above content will be affected by various conditions and variables. It will be apparent to those skilled in the art that various modifications and variations in form and detail can be made to the embodiments and implementations of this disclosure. Therefore, this disclosure also covers all such modifications, variations, and equivalents.

Claims

1. A compound of formula I or a tautomer thereof, a pharmaceutically acceptable salt, solvate, hydrate, cage compound, or polymorph thereof: I in: X is an aryl or heteroaryl group, wherein the aryl or heteroaryl group may optionally have one or more (e.g., two or three) substituents independently selected from the following: halide, cyano, nitro, C 1-6 Alkyl, -CF3, -OH, -OR 1 -NH2, -NR 2 R 3 -(C=O)NR 4 R 5 and the proteolytic targeting portion (e.g., a thalidomide-based proteolytic targeting portion), and wherein the C 1-6 Alkyl groups may optionally be replaced by -OH or -OR 1 -NH2 or -NR 2 R 3 replace; Y is a cycloalkyl, heterocyclic, aryl, or heteroaryl group, wherein the cycloalkyl, heterocyclic, aryl, or heteroaryl group may optionally have one or more (e.g., two or three) substituents independently selected from the following: halide, cyano, nitro, C 1-6 Alkyl, -CF3, -OH, -OR 1 -NH2, -NR 2 R 3 and -(C=O)NR 4 R 5 And wherein C 1-6 Alkyl groups may optionally be replaced by -OH or -OR 1 -NH2 or -NR 2 R 3 replace; R 1 It is hydrogen, C 1-6 Alkyl, -CHF2, -CF3 or C 3-6 Cycloalkyl, wherein the alkyl or cycloalkyl group may optionally be converted to -OH, C 1-4 Alkoxy, -NH2 or -NR 2 R 3 replace; R 2 and R 3 Independently hydrogen, C 1-6 Alkyl or -C(=O)-(C 1-6 Alkyl), or R 2 and R 3 And the nitrogen atoms they are attached to form 3- to 6-membered heterocycles or 5- or 6-membered heteroaromatic rings; R 4 and R 5 Independently hydrogen or C 1-6 alkyl; n is 1, 2, or 3; and The stereocenter marked with an asterisk (*) may have (S)-stereochemistry or (R)-stereochemistry or may be racemic; The conditions are: X is not an unsubstituted phenyl or 4-chlorophenyl; and The compound of formula I is not: 、 、 or , Or its tautomers or salts.

2. The compound according to claim 1, wherein X is an optionally substituted 5- to 10-membered monocyclic or bicyclic aryl or heteroaryl.

3. The compound according to claim 1 or 2, wherein X is selected from 2-CN-phenyl, 3-CN-phenyl, 3-CH3-phenyl, 3-OCH3-phenyl, 3-(C(=O)NH2)-phenyl, 4-Br-phenyl, 4-OCH3-phenyl, 4-OCHF2-phenyl, 4-(morpholino-N-yl)-phenyl, 3-CN-4-CH3-phenyl, 3-F-4-OCH3-phenyl, 3-F-5-CH3-phenyl, 3,5-Dimethyl-phenyl, 1-naphthyl, N-CH3-pyrazol-4-yl, 4-CH3-pyridin-2-yl, 5-CN-pyridin-2-yl, 5-CH3-pyridin-2-yl, 5-CF3-pyridin-2-yl, 6-CH3-pyridin-2-yl, 2-CH3-pyridin-3-yl, 4-CH3-pyridin-3-yl, 5-CH3-pyridin-3-yl, 6-CH3-pyridin-3-yl, 2-CH3-pyridin-4-yl , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

4. The compound according to any one of the preceding claims, wherein for Y: The cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, or cyclohexenyl; The heterocyclic group is tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, piperazineyl, or morpholinyl; The aryl group is phenyl, naphthyl, dihydroisobenzofuranyl, methylenedioxyphenyl, benzodioxane, isoindolinyl, 1,2,3,4-tetrahydroquinolinyl, or 1,2,3,4-tetrahydroisoquinolinyl; and The heteroaryl group is furanyl, thiophene, pyrrole, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, quinolinyl, or isoquinolinyl; The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted as defined in claim 1.

5. The compound according to claim 4, wherein Y is selected from cyclohexyl, 1,1-diF-cyclohexyl-4-yl, 2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl, piperidin-3-yl, N-CH3-piperidin-3-yl, N-acetyl-piperidin-3-yl, piperidin-4-yl, N-CH3-piperidin-4-yl, N-acetyl-piperidin-4-yl, phenyl, 2-F-phenyl, 2-Cl-phenyl, 2-CF3-phenyl, 2-OCH3-phenyl, 2-OCH2CH3-phenyl, 2-OCH2CH2CH3-phenyl, 3-F-phenyl, 3-Cl-phenyl, 3-CH3-phenyl, 3-CF3-phenyl, 3-OCH3-phenyl, 3-(OCH2CH2O H)-phenyl, 3-(piperidin-N-yl)-phenyl, 3-(morpholin-N-yl)-phenyl, 3-(imidazol-N-yl)-phenyl, 4-F-phenyl, 4-Cl-phenyl, 4-Br-phenyl, 4-CH3-phenyl, 4-ethyl-phenyl, 4-isopropyl-phenyl, 4-CH2OH-phenyl, 4-CF3-phenyl, 4-OH-phenyl, 4-OCH3-phenyl, 4-OCHF2-phenyl, 4-OCF3-phenyl, 4-(OCH2CH2OCH3)-phenyl, 4-(O-cyclopentyl)-phenyl, 4-N(CH3)2-phenyl, 4-(N-acetyl)-phenyl, 4-(imidazol-N-yl)-phenyl, 4-(pyrazol-N-yl)-phenyl, 2-CH3-3- F-phenyl, 2-OCH3-3-F-phenyl, 2-F-4-Cl-phenyl, 2,4-diCl-phenyl, 2,4-dimethyl-phenyl, 2-F-4-methoxy-phenyl, 2-F-4-ethoxy-phenyl, 2-OH-4-OCH3-phenyl, 3-F-4-Cl-phenyl, 3,4-diCl-phenyl, 3-CH3-4-F-phenyl, 3-CF3-4-CH3-phenyl, 3,5-dimethyl-phenyl, 1-naphthyl, 2-naphthyl, 1,2,3,4-tetrahydroisoquinoline-8-yl, 4-isoindolinyl, 5-dihydroisobenzofuranyl, 1,3-methylenedioxyphenyl-5-yl, 1,4-benzodioxane-6-yl, N-CH3-pyrazol-4-yl, N-CH 3-3-CH3-pyrazol-4-yl, N-methyl-imidazol-2-yl, N-ethyl-imidazol-2-yl, 5-CH3-isoxazol-3-yl, 2-pyridyl, 3-F-pyridyl-2-yl, 3-CH3-pyridyl-2-yl, 4-Cl-pyridyl-2-yl, 4-CH3-pyridyl-2-yl, 4-OCH3-pyridyl-2-yl, 5-Cl-pyridyl-2-yl, 6-CH3-pyridyl-2-yl, 4,6-dimethyl-pyridyl-2-yl, 3-pyridyl, 2-OCH3-pyridyl-3-yl, 6-OCH3-pyridyl-3-yl, 4,6-diCl-pyridyl-3-yl, 4-CH3-pyrimidin-2-yl, 5-CH3-pyrimidin-2-yl, 2-quinolinyl, and 5-isoquinolinyl.

6. The compound according to any one of the preceding claims, wherein n is 1 or 2.

7. The compound according to any one of the preceding claims, wherein it is one of the following compounds, or a tautomer, pharmaceutically acceptable salt, solvate, hydrate, cage compound, or polymorph thereof: ； The stereocenter marked with an asterisk (*) is racemic or has (S)-stereochemistry, and compounds exhibiting (S)-stereochemistry may alternatively be racemic at that stereocenter.

8. The compound according to any one of claims 1 to 6, wherein it is one of the following compounds, or a tautomer, pharmaceutically acceptable salt, solvate, hydrate, cage compound, or polymorph thereof: The stereocenter marked with an asterisk (*) is racemic or has (S)-stereochemistry, and compounds exhibiting (S)-stereochemistry may alternatively be racemic at that stereocenter.

9. A compound selected from the group consisting of tautomers, pharmaceutically acceptable salts, solvates, hydrates, cage compounds, and polymorphs thereof: The stereocenter marked with an asterisk * is racemic or has (S)-stereochemistry, and compounds exhibiting (S)-stereochemistry may alternatively be racemic at the stereocenter, and Boc represents tert-butyloxycarbonyl.

10. The compound according to any one of the preceding claims, wherein the stereocenter marked with an asterisk (*) has (S)-stereochemistry.

11. A pharmaceutical composition comprising the compound or tautomer thereof as described in any one of the preceding claims, a pharmaceutically acceptable salt, solvate, hydrate, cage compound or polymorph, and one or more pharmaceutically acceptable excipients or carriers.

12. The pharmaceutical composition according to claim 11, wherein it is an oral dosage form, such as a tablet, capsule, or pill.

13. The pharmaceutical composition according to claim 11 or 12, further comprising an additional therapeutic agent.

14. A method of treating a tumor or cancer, comprising administering to a subject an effective amount of any one of claims 1 to 10, a tautomer thereof, a pharmaceutically acceptable salt, a solvate, a hydrate, a cage compound, or a polymorph thereof, to treat the tumor or cancer.

15. The method of claim 14, wherein the tumor or cancer is selected from solid tumors, head and neck tumors and cancers (e.g., oral squamous cell carcinoma, esophageal squamous cell carcinoma, nasopharyngeal carcinoma, and papillary thyroid carcinoma), CNS tumors and cancers (e.g., glioblastoma, glioma, and neuroblastoma), lung cancer (e.g., non-small cell lung cancer), gynecological malignancies (e.g., breast cancer, ovarian cancer [e.g., high-grade serous carcinoma], and uterine cancer [e.g., cervical cancer and endometrial cancer]). Cancers of the digestive tract (e.g., stomach cancer [e.g., malignant epithelial tumors of the stomach] and colorectal cancer), liver cancer (e.g., hepatocellular carcinoma and intrahepatic bile duct cancer), pancreatic cancer (e.g., pancreatic adenocarcinoma), urinary system cancers (e.g., kidney cancer [e.g., clear cell renal cell carcinoma and papillary renal cell carcinoma], bladder cancer and prostate cancer), skin cancer (e.g., melanoma), sarcoma, and blood and lymphomas (e.g., leukemia and lymphoma [e.g., non-Hodgkin lymphoma such as diffuse large B-cell lymphoma]).

16. The method of claim 14 or 15, wherein the tumor or cancer is characterized by intraperitoneal, peritoneal, or / and omental metastases, such as ovarian cancer (e.g., epithelial ovarian cancer), endometrial cancer, gastrointestinal cancer (e.g., gastric cancer or colorectal cancer), or pancreatic cancer (e.g., pancreatic ductal adenocarcinoma).

17. The method according to any one of claims 14 to 16, wherein the compound is administered orally.

18. The method according to any one of claims 14 to 16, wherein the compound is administered intratumorally.

19. The method according to any one of claims 14 to 18, further comprising administering one or more additional antitumor / anticancer agents.

20. A method of treating metabolic disorders or liver diseases, comprising administering to a subject an effective amount of the compound or its tautomer, pharmaceutically acceptable salt, solvate, hydrate, cage compound or polymorph of any one of claims 1 to 10 to treat metabolic disorders or liver diseases.

21. The method of claim 20, wherein the metabolic disorder is selected from obesity, obesity-related disorders (e.g., cardiovascular disorders and hypertension), hyperlipidemia (e.g., hypercholesterolemia and hypertriglyceridemia), metabolic syndrome, hyperglycemia, impaired glucose tolerance, insulin resistance, type 2 diabetes, polycystic ovary syndrome (PCOS), and age-related disorders (e.g., sarcopenia).

22. The method of claim 21, wherein the metabolic disorder is obesity, obesity-related disorder, or type 2 diabetes.

23. The method of claim 20, wherein the liver disease is selected from hepatic steatosis, alcoholic liver disease (ALD), non-alcoholic fatty liver disease (NAFLD), hepatitis, alcoholic steatohepatitis (ASH), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis.

24. The method of claim 23, wherein the liver disease is ALD, NAFLD, ASH, or NASH.

25. The method according to any one of claims 20 to 24, wherein the compound is administered orally.

26. The method according to any one of claims 20 to 25, further comprising administering one or more additional therapeutic agents.

27. The method of claim 26, wherein the one or more additional therapeutic agents comprise one or more anti-obesity agents and / or one or more anti-diabetic agents.

28. A method for inhibiting nicotinamide N-methyltransferase (NNMT), comprising contacting cells with the compound or tautomer of any one of claims 1 to 10, a pharmaceutically acceptable salt, solvate, hydrate, cage compound or polymorph.

29. The method of claim 28, wherein the contact occurs in vivo, outside vivo, or in vitro.

30. A method for regulating metabolites in the NAD+, methionine, and S-adenosyl-L-methionine (SAM) / S-adenosyl-L-homocysteine ​​(SAH) pathways using the compounds of any one of claims 1 to 10.

31. A method for regulating gene expression or epigenome using the compound of any one of claims 1 to 10.

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